Title of Invention

A METHOD FOR COLOSTRAL FRACTIONATION

Abstract A method for separating the proline-rich polypeptides essentially free of peptides and proteins of about 10,000 Daltons or greater from the globular proteins having a molecular weight of between 10,000 and 500,000 Daltons in bovine colostrum, the method comprising the steps of: (a) subjecting at least a portion of a bovine colostrum sample to at least one separation procedure comprising an ion exchange separation to yield separated proline-rich polypeptides essentially free of peptides and proteins of about 10,000 Daltons or greater and separated globular proteins having a molecular weight of between 10,000 and 500,000 Daltons, wherein no more than about 10% of the separated proline-rich polypeptides are denatured; and (b) collecting the separated proline-rich polypeptides; wherein at least 90% of the separated proline-rich polypeptides have a molecular weight of no more than about 2,500 Daltons.
Full Text A NOVEL COLOSTRAL FRACTIONATION PROCESS
BACKGROUND OF THE INVENTION
Cross References to Related Applications
[0001] This application claims benefit of United States provisional patent application
serial number 60/516,674, filed November 3, 2003, which is herein incorporated by
reference.
Statement of the Technical Field
[0002] The present invention relates to nutritional compositions, and more
particularly to nutritional compositions that are derived from colostrums of animals.
Description of the Related Art
[0003] Since the early 1970's whey proteins have been concentrated for uses in
healthcare and for use in nutritional and functional foods, for example as an
ingredient. These proteins include beta-lactoglobulin and alpha-lactalbumin, which
are commonly extracted from bovine milk. More recently, lactoferrin and
immunoglobulins also have been extracted from bovine milk, but to a lesser extent.
Although the concentrations of lactoferrin (100-200 mg/liter) and the
immunoglobulins (5-6% of total whey proteins) are low in milk, the concentrations of
lactoferrin (1-2 g/liter), proline-rich polypeptides and immunoglobulins (70-80% of
total whey proteins) are much higher in colostrum.
[0004] Some protein functions involve the binding of other molecules called ligands.
Ligands can be drugs, hormones or antigens that can bind with proteins. Those
compounds that can act as ligands but which normally are found naturally in animal
bodies fall into three general classes: neurotransmitters, steroids (including the sex
hormones), and peptides. The first two classes are considered to be bioactive,
whereas peptides are not. (Bioactive is a phrase that describes a set of compounds
which have an effect on animal cells that is in direct proportion to their

number and which are produced either outside the body of the animal or only in
specialized organs or systems thereof.) Peptides typically have a relatively low
molecular weight and are the product of the sequential covalent bonding of several
amino acids. For example, peptides typically are formed from about 4 to 100
sequentially bonded amino acids.
[0005] Proline-rich-peptides and/or non-ionic peptides are important in various
biochemical processes. These peptides may be termed receptor peptides and are in
fact very specialized types of proteins. They are believed to reside in or on the
exterior surfaces of all animal cells (regardless of particular cellular function). When
activated through interaction with a ligand, a receptor then transmits a biochemical
message into the interior of the cell.
[0006] Peptides, such as proline-rich-peptides and non-ionic peptides, which
function as ligands are produced in the ribosomes of all or very nearly all of the cells
of animals and man. They are sometimes referred to as informational peptides
because they often provide little or no nutritional value, but contain specific
information to help trigger specific biological processes. The informational peptides
also may help protect cells by re-orientating receptor sites often used by synthetic
viral protein ligands. Thus, these peptides may help inhibit viruses from attaching
themselves to those specific individual target cells by regulating immuno-modular
and cytokine intercellular function and intracellular function.
[0007] Peptides generally are relatively small, at least in relation to most proteins
which tend to have molecular weights of at least 20,000 Daltons. In general,
peptides have a molecular weight of no more than about 1000 Daltons, although
some might be larger, even perhaps as large as about 6000 Daltons. Nevertheless,
they are significantly smaller than most proteins.
[0008] When a peptide leaves the cell in which it was produced, it moves
throughout the body by way of the interstitial fluids between the cells and the
circulatory system. In the blood and interstitial fluids, peptides tend not to
agglomerate with themselves (i.e., they remain separate). This separateness allows

the peptides to remain in forms in which they can bind with appropriate receptors.
For instance, a peptide produced by one cell can be transported to and interact with
the cellular function of a distant cell. When such an interaction occurs, a type of
biochemical transmission to the cell interior is set into motion and this, in turn,
induces some type of a response within the cell. One such cellular action is believed
to be the production of additional peptides of the type bound to the cellular receptor.
[0009] As mentioned previously, some viruses take up residence in animal bodies
by entering cells through particular types of receptors. If the necessary type of
receptor already is bound to another ligand, such as a peptide, or the shape of the
receptor does not or is no longer compatible with the viral ligand, then the virus
cannot enter that given cell and must find another cell in which to enter. If all cells
have the target receptor bound with other ligands, or there has been a
conformational change of shape at the receptor site because of biochemical
processes from within the cell, the virus' entry path is blocked and infection is
averted.
[0010] When an animal, including a human, is healthy, it has a full (or very nearly
full) complement of peptides. However, due to any one or more of a variety of
factors, such as increased age of the animal, bodily abuse by environment or
substance abuse, nutrition, suppressed immune system, and/or illnesses and
diseases, an animal may fail to produce or maintain one or more of these types of
peptides. Such failures often can be the first cause of illness. Return to health can
be relatively quick and easy, however, when the missing peptide(s) is reintroduced
into the body because such peptides can, as described above, "instruct" cells to
create more copies of the peptides. These are commonly called "proline-rich-
polypeptides" (PRPs), "cytokine precursors" or "immuno-modulating peptides".
Commonly, these peptides have been called the "software of the cell" or "software of
the human operating system", which refer to the information required for all living
mammalian cells to function. The initialization of correct cellular function is started
when a female lactating mammal first delivers the "colostrum" to a newborn mammal
baby, which commonly is called "passive immunity". In addition to such immunity,
the colostrum also provides cytokine precursors to initiate many biochemical

processes in mammalian cells. Thus, reintroduction of a small amount -perhaps a
single copy- of one or more missing peptides to any infant, teenage, adult or elderly
human, or any aged mammal, can quickly return cells in the body to their normal
amount of the peptide(s) in question.
[0011] The target peptides can be derived from blood or from other mammalian
bodily fluids derived from or in contact with blood. Such fluids include, but are not
limited to, milk, colostrum, semen, urine, vaginal fluid, and the like. However, in
materials such as milk and colostrum, for example, peptides are in what is
essentially an impaired state because they are agglomerated with or on much larger
biochemical macromolecules i.e. fats or other proteins. Additionally, ingestion by
eating or drinking certainly denatures the peptides because of the acidic conditions
of the stomach and the relatively aggressive enzymatic action of the digestive tract.
Thus, although many external sources of peptide ligands are available, these
peptides often are in a form that renders them useless for the desired effect.
Accordingly, processing or refinement of such external sources is necessary.
[0012] Of the external sources of peptides, the one that seems to provide them in
the highest concentrations and is most widely available is colostrum. This material
has been the subject of numerous processing methodologies. However, almost all of
the previously described processing methods appear to have been directed at
collecting or isolating biologically active macromolecules that are much larger than
peptides, such as, for example, proteins, lactoferrin, immunoglobulin, lipids, etc.
[0013] Importantly, present colostrum processing methods tend to encourage
relatively high fluid pressures and much lower yields of peptides as a side effect of
fast processing speeds and current technologies used. For those references dealing
with ways to isolate large molecules such as immunoglobulin, lactoferrin, etc., this is
not surprising because such macromolecules are relatively hearty and capable of
withstanding such pressures. Peptides, however, respond quite differently to high
processing pressures. In particular, many types of peptides can be denatured at
pressures ranging from about 210 kPa (approximately 30 psi) to about 690 kPa
(approximately 100 psi). For example, peptides involved in the prevention of viral

infections are among those that can be denatured at the lower end of this range of
pressures (less than 10 psi). The term "Denatured", with respect to a peptide,
connotes an alteration or conformation change from the natural state due to, for
example, physical forces (e.g., adhesion to another molecule(s), exposure to
excessive temperature or pressure during processing, etc.), chemical reaction (e.g.,
scission due to exposure to excessively acidic or basic conditions), enzymatic
degradation, and the like.
[0014] Accordingly, there remains a need for a method of processing animal-
derived fluids that result in an end product which is peptide-rich, with the same
efficacy as in its native state, but substantially free from other materials that can
denature such peptides, and thus able to fully express their peptide bioactivity
without steric hindrance, and increase liquid diffusion of these peptides.
SUMMARY OF THE INVENTION
[0015] The present invention relates to a method and a system for processing
mammalian bodily fluids, for example colostrum, to isolate target peptides and
proteins. The mammal can be a bovine, for example a cow, goat, pig, buffalo, deer,
or any other suitable mammal. For example, the present invention can process
colostrum produced by lactating cows, but is not so limited. Notably, other mammals
can be used to supply colostrum, or other mammalian fluids.
[0016] The method can include the steps of passing a mammalian bodily fluid
through at least one ion exchange column or filter. The ion exchange column can
include an anionic resin and/or a cationic resin. The ion exchange column or filter
can be selected to remove large particles, for instance certain proteins, at a first pH
level selected to remove such large particles at a maximum fluid pressure of 30
pounds per square inch (psi). In another arrangement, the maximum fluid pressure
can be less than 10 psi. The large particles can be released from the ion exchange
column or filter with a rinse solution having a pH selected for releasing the large
particles.

[0017] The pH of the fluid then can be adjusted to be a value less than 5.0, and
the fluid can be filtered with a microfilter having an initial pore size no greater than
300 nm or an ultrafilter having a pore size no greater than 20,000 Daltons. A pH of a
retentate trapped by the filter can be adjusted to have a value between 6.5 and 7.0
and antibiotics can be washed from the retentate.
BRIEF DESCRIPTION OF THE DRAWINGS
[0018] FIG. 1 is a flow chart of a process which is useful for understanding the
present invention.
[0019] FIG. 2 is a block diagram of a processing system which is useful for
understanding the present invention.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0020] The present invention relates to a method of isolating certain peptides and
proteins from mammalian bodily fluids. In particular, the peptides and proteins can
be isolated from the bodily fluids by filtering the bodily fluids using one or more filters
having a relatively small average pore size, for example an average pore size of 1.4
urn or less. In one arrangement, the peptides and proteins being isolated can
encounter a pressure that is equal to or less than 30 pounds per square inch (psi),
and more preferably a pressure that is less than 10 psi. The peptides targeted for
isolation can be, for example, proline-rich-peptides and/or non-ionic peptides. Such
peptides are important in various biochemical processes. The proteins targeted for
isolation can be, for example, alpha-lactalbumin, beta-lactoglobulin, lactoferrin,
lactoperoxidase, IgS, and immunoglobulins such as IgG, IgA, and IgM, etc.
[0021] Blood and mammalian bodily fluids derived from blood, or in contact with
blood, are potential sources for the target peptides and proteins. For example,
colostrum, milk, semen, urine, vaginal fluid, and the like can be used as a source of
the target peptides and proteins. Advantageously, colostrum has relatively high
levels of lactoferrin, immunoglobulins, and peptides, and colostrum is produced in
relatively large quantities. Thus, for purposes of the following description of the

isolation process, colostrum is used as a representative starting material. It should
be noted, however, that the present invention is not so limited in this regard.
[0022] FIG. 1 presents a method 100 which is useful for understanding a method
for processing colostrum. The first colostrum withdrawn from a given mammal
usually has a larger amount of the target proteins and peptides (per unit volume)
than any subsequent colostrum collected from the same mammal. Thus, the first
and second milking of a particular mammal can provide a higher yield of the desired
peptides in comparison to subsequent milkings. Although the production of the
target peptides and proteins is not affected by the use of dairy cows which have had
their utters specially treated with antibiotics or antigen-like materials that have more
than a 30 day withholding period, the use of such dairy cows is not preferred
because of potential contamination of the colostrum with antibiotics. Nonetheless, if
such dairy cows are used, the antibiotics or materials can be removed from the
colostrum using filtration technologies.
[0023] Referring to step 102, the colostrum can be pre-processed. For instance,
the colostrum can be collected and stored under suitable storage conditions.
Specifically, the colostrum preferably is collected under conditions that prevent gross
contamination by bacteria. For example, the collection and storage conditions can
be those which are appropriate for the storage of milk for human consumption. For
instance, the conditions can be consistent with applicable governmental (e.g.,
USDA) guidelines.
[0024] Where pumps are employed in a given step, they preferably are of a type
that does not have impellers or other features capable of producing a shearing
effect. In the design of a specific process, each aspect preferably is considered and
tailored to prevent damage to the target peptides and proteins. Once the raw
colostrum farm tank has been emptied the tank can be cleaned with an automatic
CIP system through a spray ball. The use of a tank that does not have sharp
corners can be used to facilitate cleaning of the tank and minimize build up of
colostrum in the corners of the tank.

[0025] During processing, it is preferred that the fluid pressures which are
experienced by the target peptides at each part of the process stay equal to, or
below, 30 psi. For example, keeping the fluid pressures experienced by the target
peptides at or below about 10 psi can minimize denaturing of the target peptides.
To provide these relatively low operating pressures while maintaining industrially
acceptable processing speeds, running the process in the form of a batch (as
opposed to continuously) can be preferable. Where batch processing is used,
typical amounts can range from about 375 to about 37,500 L (100 to 10,000 gallons),
depending on down-line processing speed.
[0026] There can be a further benefit by immediately placing the raw material
under refrigeration. For example, cooling the raw material to less than 45 degrees
Fahrenheit in less than 4 hours can reduce bacterial growth in colostrum. Any
suitable cooling technique can be used to cool the raw material. For example, inline
plate heat exchangers or chilled water systems can be used to immediately cool the
colostrum to a desired temperature. Still, the invention is not so limited and other
cooling methods can be used.
[0027] Freezing of colostrum, as well as other types of raw material, reduces the
efficacy or concentration of the natural peptides and is therefore not desired because
many of the peptides and globular proteins can be damaged by the freezing of the
water within the raw material. In particular, water crystals formed when the
colostrum is frozen can cut through the proteins, and thus increase the degree of
denaturing. This can reduce the yield of target peptides and proteins from the raw
material. Nevertheless, although the use of previously frozen colostrum is not
optimal, it still can be used and is within the intended scope of the present invention.
[0028] Processing of the colostrum can be performed proximate to or remote frem
the point of collection. In the case that processing is performed remote to the point
of collection, transportation of the raw material preferably occurs under conditions
suitable for handling of the raw material without significant denaturing of the target
peptides and proteins and without excessive bacterial growth. For example, the raw
material can be transported using methods known to those skilled in the art.

[0029] Testing can be performed on the colostrum to measure the concentration
of target proteins and/or target peptides within the raw material. For example, when
attempting to collect colostrum from a mammal, an amount of milk may also be
collected. Since milk can have a lower concentration of the target peptides and
proteins than colostrum, greater milk content within the raw material can lower the
yield of the target peptides and proteins from the raw material. Specific gravity
testing can be used to measure the amount of milk present in the raw material, and
thus provide an indication of the quality of the raw material for the intended purpose.
Other methods also can be used to indicate the quality of the raw material. For
example, high-pressure-liquid-chromatography, radial immunodiffusion (RID) assay,
and/or enzyme-linked immunosorbent assay (ELISA) can be used to accurately
determine the levels of immunoglobulins present in the raw material. The testing of
the raw material can be implemented as part of a quality control program.
[0030] The amount of colostrum collected from a cow can be limited to no more
than approximately 15 L (4 gallons), collected within the first 24 hours after birth. In
the case that the colostrum is transported to a processing facility remotely located
from the location of the colostrum collection, it can be advantageous to maintain the
colostrum below about 45 degrees Fahrenheit (7 degrees Centigrade) at all times to
prevent excessive bacteria growth, but it is preferred that freezing of the colostrum
be avoided to prevent denaturing of the proteins and peptides. Further, it also can
be advantageous to process the colostrum soon after collection to insure a high
quality product.
[0031] For example, it is preferred that the colostrum arrive at the processing
facility within 72 hours of collection. The colostrum can be pasteurized using an
automated legal high temperature short residence time (HTST) pasteurizing system,
or ultra-high temperature (UHT) pasteurizing system, but preferably not batch
pasteurized. The raw material can be stored in a suitable storage container for
processing. For example, the storage container can be a tank made of a relatively
inert material, such as stainless steel.

[0032] The process also can isolate colostral whey or colostral sera from the
colostrum for further processing. Colostral whey can be derived from separation
technologies from any pre-curing of the colostrum. Colostral sera can be derived
from separation technologies from any non-curing processes of the colostrum.
When colostrum is collected from more than one mammal, the colostrum can be
stirred with some type of stirring mechanism, for example a paddle or stirrer blades,
or a pumping motion, to gently mix the raw material. Where stirrer blades are used
as the stirring mechanism in the preceding step, slow rotation of the blades and a
backward angling of the blades can provide a gentle mixing to minimize damage to
peptides and proteins. Where pumping is used as the stirring mechanism in the
preceding step, slow rotation of the impellers with low shear impeller designs can
provide a gentle mixing to minimize damage to peptides and proteins.
[0033] Such mixing can help to counter inconsistencies between colostrum from
the different mammals and to provide a more uniform temperature. For example,
because a particular cow might be deficient in one or more particular peptides, a
blending of colostrum from many cows can provide a given colostral blend that
contains all of the target peptides and proteins. Further, both cow and heifer
colostrum can be mixed together. Colostrum containing blood of non-genetically
modified mammals typically does not have a high concentration of the target
peptides, however.
[0034] After blending, the colostrum blend can be moved to a first phase of a
reduction process which involves separating and removing much fat from the
colostrum. Various means, such as separators, centrifuges, chemical, hydrophobic,
supercritical CO2 or liquid-liquid extractions are available to accomplish this task.
[0035] The defatted blend can be conveyed to a curdling vessel (e.g., a stainless
steel tank) where it can be gently stirred. During this process the temperature of the
defatted blend can be raised to between about 90 degrees Fahrenheit (32 degrees
Centigrade) and about 99 degrees Fahrenheit (37 degrees Centigrade) in
preparation for curdling. Curdling involves the coagulation of the majority of solids
remaining in the blend (principally casein). It generally is accomplished by addition of

rennin, an enzyme-rich extract from the stomachs of calves, or an acid such as HCI
to the warmed colostrum blend. Once the rennin or acid is added, a curd gradually
begins to form soon after stirring of the blend is stopped. As the curd forms, it rises
to the surface, producing a soft white cake or crust sitting or floating on whey. The
whey, which is the desired product from this step, can be drained away from the
curdling vessel. Alternatively, a defatted colostrum stream can be microfiltered
and/or ultrafiltered instead of curding to extract the same peptides. .
[0036] The remaining curd can be cut into small pieces by, for example, activating
a stirring mechanism in the tank. The broken curd can be conveyed away from the
tank through pipes made of an inert material, for example stainless steel, to a large
screen, which also can be made from a relatively inert material. Any whey trapped in
the curd can pass through the screen and, optionally, can be added into the whey
collected previously. (The curd can be collected for sale or discarded, as desired.)
[0037] Proceeding to step 104 of FIG. 1, the collected whey or sera can be
passed through a fines reducer or a clarifier to exclude more of the small pieces of
curd which may be conveyed away from the curdling vessel during removal of the
whey. This additional step, although certainly not required, can be beneficial
because it increases the service interval for the filter media described below. The
whey can be conveyed to the next step of the reduction process through, for
example, stainless steel pipes.
[0038] Making reference to the processing system 200 of FIG. 2, the pre-
processed colostral whey (or colostral sera) 202 can undergo further processing
using ion exchange chromatography to isolate desired proteins from the colostral
whey 202. Proceeding to step 106 of FIG. 1, the pH of the colostral whey 202 can
be adjusted as required for large protein sorption by one or more ion exchange
columns 204, which can be a cationic ion exchange column or an anionic ion
exchange column. The ion exchange column 204 can comprise resin, which can be
in the form of a bead of uniform size or with a wide standard deviation of sizes, or a
non-spherical shape of similar or varying sizes. For example, a cationic resin can be
an SP or SM type resin, and an anionic resin can be a QEA or Q type resin. Such

resins are known to those of ordinary skill in the art. The resin can be packed with
adequate bulk density or contained within a containment vessel so as to allow
adequate flow of the colostral whey 202.
[0039] A pH adjustment solution 206 can be added to the colostral whey 202 until
a suitable pH of the colostral whey 202 is reached. Notably, the pH of the colostral
whey 202 can affect the surface chemistry of proteins contained within the colostral
whey 202, and give the surface of the proteins a net negative charge, a net positive
charge, or a net neutral charge. A suitable acid for lowering the pH of the colostral
whey 202 can be, for example, food grade citric acid, phosphoric acid, or lactic acid.
Alternatively, a suitable base can be added to the whey to raise the pH. Examples of
suitable bases are potassium hydroxide, calcium hydroxide and sodium hydroxide,
but the present invention is not limited in this regard. Notably, the.use of potassium
hydroxide adds potassium to the colostrum, which protects proteins during a
subsequent pasteurization process, if such a process is used. A suitable valve 214
and/or suitable mixing pumps (not shown) can be used to mix the pH adjustment
solution 206 with the colostral whey 202.
[0040] Continuing at step 108, the colostral whey 202 can be passed through the
ion exchange column 204 at a pressure less than or equal to 30 psi, and more
preferably below 10 psi, to isolate one or more globular proteins between 10,000 and
500,000 Daltons. For example, such proteins can include alpha-lactalbumin, beta-
lactoglobulin, lactoferrin, immunoglobulins and other whey proteins. A cationic ion
exchange column can comprise a positive resin. In consequence, lactoferrin arid
lactoperoxidase can be sorbed when the pH of the whey solution is between about
6.5 - 7.0. Moreover, all whey proteins can be sorbed within the cationic ionic
exchange column when the pH is less than 4.0. For instance, the pH can be in the
range of about 3.0 to 4.0. An anionic ion exchange column can comprise a negative
resin. In this arrangement, all whey proteins can be sorbed when the pH of the whey
solution is between about 6.5 - 7.0. Typically, little or no proteins are sorbed in an
anionic exchange column when the pH of the solution is less than 4.0.

[0041] Proteins sorbed in the resin are globular proteins that typically do not bind
to the resin. Since the proteins do not bind to the resin, the proteins can be
recovered from the ion exchange unit 204 by passing a rinse solution 216 having a
pH selected to release the proteins, as shown in step 110. In particular, the rinse
solution 216 can change the charge of the resin in the ion exchange column 204 and
cause the resin to release the proteins into a retentate stream. Solutions also can be
used to change the net charge of the surface of the proteins so that the proteins
repel the cationic or anionic resins. Proceeding to step 112, the retentate stream
with the liberated proteins then can be difiltered to remove any salts that may remain
in the solution. For example, the retentate stream can be passed through one or
more filters 218 which have pore sizes greater than the globular proteins in the
retentate stream. The retentate stream then can be dried to recover the globular
proteins, as shown in step 114. For instance, the proteins can be concentrated to
between 15-30% total solids. The proteins then can be passed through a dryer 220
and subjected to freeze or low-heat indirect steam spray drying. An evaporator can
be used to concentrate further the retentate stream. Suitable valves 214 and/or
suitable fluid pumps (not shown) can be used to control fluid flow within the system
200.
[0042] Advantageously, the whey or sera need not be subjected to fluid pressures
in excess of normal bodily fluid pressures in this process, thus preserving the native
states of the proteins and peptides contained in the whey. The resulting filtrate or
flow-through stream therefore can be more effective in cellular interactions in
comparison to a filtrate filtered at pressures which are significantly higher than the
pressures the bodily fluids experience within the mammal from which they are
extracted. Also, the yield of non-ionic peptides from the whey or sera using ion
exchange technologies, as will be described below, can be between 80-85% when
compared to the original number in the raw colostrum liquid. When membrane
filtration techniques are exclusively used to isolate the proteins, a peptide yield of
about 45-50% can be expected. This is a significant yield improvement over existing
extraction technologies.

[0043] In the foregoing step, to achieve adequate separation while maintaining
operating pressures at or below about 30 psi, for example below about 10 psi,
relatively short filtration units can be used to minimize back pressure. For example,
the filtration units can be on the order of about 0.3 m to 1.5 m in length ion exchange
columns with fixed beds, packed beds or expanded bed designs, having bed depths
not exceeding 20 cm to 25 cm, thus minimizing the pressure drop across the bed,
preferably less than 5 psi. Notably, dynamic or moving ion exchange beds do not
have such pressure drops. However, the yield can be less with dynamic beds as
compared to static or fixed beds. Specifically, there can be a higher concentration of
proteins in the whey using dynamic beds as compared to exposing the stream of
whey to fresh resin in lower sections of a fixed bed.
[0044] In one arrangement, two or more ion exchange columns can be cascaded
in series. The use of cascaded ion exchange columns provides a longer contact
time for the flow-through protein solution. Moreover, multiple ion exchange columns
can be connected in parallel to increase throughput and scalability of a commercial
ion exchange process. Ion separation technologies as described herein can be used
before or after the filtration steps as needed to purify individual or multiple protein
streams, depending on the order in which it is desired to remove the proteins.
Notably, the order of ion exchange and filtration does not affect production of desired
peptides.
[0045] Although not critical to the present process, the diameter of the fixed ion
exchange beds can be minimized, for example to less than 30 cm, to prevent uneven
bed depths of resin across the resin bed to minimize "uneven lateral resin migration".
Minimizing the lateral resin migration can help to maintain a uniform pressure drop
through the ion exchange bed.
[0046] In lieu of ion exchange columns 204, tangential flow (sometimes called
cross-flow) filters can be used; tubular microfilter unit also can be used. Tangential or
cross flow filtration is generally preferred over dead-end filtration because the latter
can result in unacceptable pressures unless throughput speeds and volumes are
kept quite low. Additionally, tangential flow units do not result in all particles being

trapped in the filter membrane, i.e., certain large particles merely pass along the
exterior of the membrane and never get retained in a pore; this extends the
operation period for a given filter unit.
[0047] Microfilters as a class generally are used to remove substances that range
in size from about 0.1 to about 1.4 µm. By selecting a filter with an average pore size
of from 0.1 to 0.45 µm, one can achieve the desired result of removing most
materials having a molecular weight of approximately 500,000 Daltons or more,
which includes almost all bacteria but very few proteins. In fact, proteins the size of
antibodies and smaller pass through this sjze of filter pore with the greater portion of
the water present in the whey.
[0048] Microfilter membranes can be made from a wide variety of materials and
are commercially available from numerous sources. Membranes can have inner
permeate tube diameters of about 0.05 inch to 2.0 inches and outer diameters of
about 1.0 inch to 12.0 inches. Acceptable microfilters for such units include, for
example, ceramic filters, metallic, Teflon, polyethylsulphone (PES) (other polymeric
spiral wound), tubular, poly-plastic membranes, etc.
[0049] The colostral whey (or colostral sera) 202 can be conveyed, again
preferably by inert means such as stainless steel pipes, to the next step in the
reduction process. Here, suspended materials that range in size down to that which
can be seen only with a relatively powerful microscope can be removed, for example
those particles greater in size than 100,000 Daltons. Referring to step 116, the pH of
colostral whey 202 again can be adjusted with a pH adjustment solution 208. For
instance, a suitable base or suitable acid can be added to the colostral whey 202
until the pH is in the range from about 4.5 to about 5.0. Although not required, it is
preferable that the pH be in the range from about 4.55 to about 4.70, and most
preferably of about 4.6. This slightly acidic pH has been found to be sufficient to kill
or disable most bacterium that might have made its way through the processing or
that might have been in the container used to hold the filtrate product. The need for
preservatives and anti-bacterial, yeast and mold inhibitors can therefore be reduced
or eliminated.

[0050] Continuing at step 118, the twice pH adjusted colostral whey 202 can be
passed through microfilter(s) and/or ultrafilter(s) 210 to remove macroscopic
particles of approximately 20,000 Daltons or more while maintaining the pressure to
which the remaining proteins and peptides in the colostral whey are exposed to at
less than 30 psi. For instance, an exemplary separation process which implements
a tangential flow unit having an ultrafilter membrane. Nanofiltration or reverse
osmosis also can be used. However, nanofiltration and reverse osmosis can be
expensive and typically require higher operating pressures than ultrafiltration. In
either case, tangential flow filtration can result in lower operating pressure than
dead-end filtration.
[0051] A tangential flow ultrafilter unit can be similar in design to the previously
described microfilter unit, except that its filter can have smaller pores. For example,
the average pore size of this type of filter can range from about 1,000 to about
100,000 Daltons. An ultrafilter for such a unit can include, for example, a spiral
wound filter (Kock; Osmonics-Desal, Synder, PTI, Pall). The membranes of the filter
can be made of cellulosic materials, fluoropolymers, polysulfones, or any other
suitable material.
[0052] In some instances, the acidic pH of the liquid will cause the pore sizes of
polymer or non-rigid plastic filter membranes to shrink. For instance, a pH of 4.6 can
cause the pores of certain microfilter membranes having an initial pore size greater
than 120,000 Daltons to shrink to 50,000 Daltons, or smaller. For example, the
average pore size can be in the range from 300 nm to 450 nm (0.3 urn to 0.45 urn).
A preferred average pore size after shrinkage is from 15,000 to about 30,000
Daltons, with 20,000 Daltons being a highly preferred average pore size. Use of
such relatively small pores with acidic pHs, as described, results in the removal of
most proteins (including antibodies) and endotoxins, which might have been included
in the original colostral blend. More particularly, transmission of the antibiotic
residues through the membrane is inhibited, and the residues thus remain in the
retentate. In consequence, the acidic permeate that is produced is free from
antibiotic residues and thus can be used as a good source of proline-rich -
polypeptides.

[0053] In the two preceding reduction (i.e., filtration) steps, the temperature of the
ion exchange columns and filtration units can be maintained as high as about 52
degrees Centigrade, primarily to control bacterial growth. Alternatively, process
temperatures for all unit operations can be maintained to be no higher than 0.50 to 5
degrees Centigrade. Use of such relatively low temperatures has been found to keep
any fat remaining in the whey in macroscopic globules that do not even enter, and
thus occlude, the pores of the filters. The low temperature also maintains low growth
of microbes in the process lines and fluids. Nevertheless, the invention is not limited
in this regard and any other suitable temperatures can be used. The antibiotic free
permeates then can be collected as products and set aside from further Ion
exchange separations.
[0054] Where the ultrafiltration step employs a filter having an average pore size
of no more than about 20,000 Daltons, the filtrate product is essentially free of
proteins, peptides, and biochemical macromolecules that have a molecular weight of
20,000 Daltons or greater. For example, antibiotics, such as penicillin G, will be
removed from the filtrate product during the ultrafiltration step. More preferably, the
filtrate product is essentially free of peptides or proteins having molecular weights of
about 10,000 Daltons or greater, and most preferably at least 90% of the component
peptides of the filtrate product have molecular weights of no more than about 2,500
Daltons. This is important for several reasons, one of which is that the filtrate then is
substantially free of molecules that can act as points of agglomeration for the desired
peptides. A peptide that has agglomerated to other peptides or to larger proteins
usually is denatured. Similarly, peptides subject to bacterial-induced enzymatic
degradation also can be denatured and, accordingly, the filtrate product preferably is
essentially free of such bacteria. Moreover, the filtrate product preferably has no
more than about 10% of its component peptides in a denatured state. Even lower
concentrations of denatured peptides are desirable.
[0055] Proceeding to step 120, this antibiotic free low pH filtered permeate (filtrate
product) can be collected in a permeate tank 212 until the flow rate drops
significantly and used for further processing as liquid or powder colostrum products.
The resulting antibiotic free low pH permeate (filtrate product) which passes through

the microfilters and/or ultra filters can be collected until the flow rate drops
significantly and used for further processing as liquid or powder colostrum products.
[0056] Although not essential, the filtrate product can be further preserved by
adding food grade 0.1% EDTA to inhibit bacterial growth and 0.1% Potassium
Sorbate to minimize any yeast and mold from growing in the final product, or
hydrogen peroxide, or some other suitable preservative. If desired, the filtrate
product may have the flavor profile changed by adding a flavor ingredient such as
vanilla, chocolate, strawberry, pina-colada, etc. This may make the filtrate product
more palatable as an ingested liquid, spray, capsule, or lozenge. If the filtrate is to
be used as topical liquid or topical spray only, scent could be added to the filtrate.
As a final treatment the filtrate product can be further filtered one or more times
using a filter not more than 0.45 urn to catch any potential remaining microbes. The
filtrate then can be aseptically packaged. At all stages of the processes Good
Manufacturing Practices (GMPs) should be followed to preserve the integrity of the
final filtrate product.
[0057] Each milliliter of filtrate product preferably includes at least 0.3 g of total
protein, more preferably at least 0.35 g protein. Most preferably, each milliliter of
product contains from 0.36 to 0.38 g of peptides. Although this amount seems small,
it has been found to provide a full (or very nearly full) array of target peptides, thus
providing maximum benefit to the human or animal which receives a dose thereof.
[0058] At this point it should be noted that the ion exchange separation
technologies as described herein can be used before or after the filtration steps.
Indeed, the ion exchange and filtration steps can be implemented as required to
achieve desired protein streams, depending on the order one wishes to remove
these proteins. However, the process technology order is irrelevant with respect to
the final filtrate product because the desired peptides are not affected by the order in
which the colostral whey 202 is processed by the ion exchange columns 204 Or
filters 210.

[0059] In the present example, the retentate next can be collected from the
microfilters/ultrafilters. At step 122, the pH of the retentate collected in the
microfilters and/or ultra filters can be adjusted to be above 5.0, and preferably in the
range 6.5 to 7.0. The pH can be adjusted using a basic solution. For example, a
typical commercial food grade calcium hydroxide, potassium hydroxide and/or
sodium ammonium hydroxide can be used to adjust the pH of the retentate. At step
124, the retentate can be released from the microfilter(s) and/or ultrafilter(s) 210 with
a rinse solution 222 to recover filtrate product from the filters 210. The rinse solution
222 can be a salt solution or other suitable permeate stream with some ionic
strength to wash antibiotics from the retentate. Proceeding to step 126, the retentate
stream with the liberated filtrate product then can be difiltered to remove any salts
that may remain in the solution. For example, the retentate stream can be passed
through one or more filters 224 which have pore sizes greater than the filtrate
product in the retentate stream.
[0060] Experiments have shown that washing the retentate with an ionic solution
at neutral pH and three times the volume of the retentate was sufficient to reduce the
antibiotic levels to less than 5 ppb for all types of penicillins, which is below the
United States Department of Agriculture (USDA) maximum level for acceptable
milks. Moreover, the rinse solution 222 circulated to repeat the washing cycle, which
can help to further reduce the antibiotic levels. In one arrangement, the rinse
solution 222 can be a waste stream from another dairy process. The wash/difilter
process can be repeated until the antibiotics reach a low enough level. The rinse
stream which passes through the microfilters and/or ultra filters can be collected until
the flow rate drops significantly, and discarded as a waste product.
[0061] The retentate then can be dried to recover the filtrate product, as shown in
step 126. For instance, the filtrate product can be passed through a dryer 226 and
subjected to freeze or low-heat indirect steam spray drying. An evaporator can be
used to concentrate further the filtrate product. Again, suitable valves 214 and/or
suitable fluid pumps (not shown) can be used to control the fluid flow. All stages in
the process can adhere to a Hazard-Analysis and Critical-Control Program (HACCP)

and Good.Manufacturing Processes (GMPs) consistent with the recommended
procedures of the USDA and Food and Drug Administration (FDA).
[0062] The mammal which produces the raw material used for processing can be
a mammal which has immunity to an antigen. For example, the mammals can be
genetically engineered. Nonetheless, the invention is not limited in this regard as the
mammal can be non-genetically engineered. The mammal can be rendered immune
by injecting an antigen or a protein specific for the antigen into the mammal. In
another arrangement, the mammal can be exposed to the antigen before the
mammal commences lactation. The antigen can be a virus or can be derived
therefrom. For example, the virus can be the Human Immunodeficiency Virus (HIV,
type 1 or type 2). The virus also can be a planar warts virus, an influenza virus, a
cold virus, or any other virus.
[0063] Exposure by the mammal producing the raw material to the virus can
result in peptides and proteins being produced which can be used to fight ailments.
Accordingly, the filtrate product can be administered to maintain wellness or to
induce recovery from a wide range of infectious and progressive disease processes.
For example, the filtrate product can be used to treat Allergies, Arthritis, Benign
Prostatic Hyperplasia (such as the inflammatory aspect), Cancer (for example for
adjunctive use), Celiac Sprue, Crohn's Disease, Diabetes Type II, Hypertension,
Lupus (Discoid and Systemic), Multiple Sclerosis, Perthes Disease (Active),
Premenstrual Syndrome and Endometriosis, Prion Disease (Kuru and Creutzfeld-
Jakob Syndrome), Psoriasis, Sjogren's Syndrome, Spinal Muscular Atrophy,
Thrombocytopenia (Ideopathic and Autoimmune), Topical Applications (burns,
wounds, infections, insect bites, diaper rash and Herpetic Lesions), Acute Viral
Infections, and numerous other ailments. Further, peptides and proteins isolated
from the raw material can be used to treat digestive problems. Still, there are a
number of other medical uses for such proteins, and the invention is not so limited.
[0064] The above list of conditions and diseases are simply a sample of what can
be treated indirectly by a course of peptide treatment, and is by no means limiting.
Moreover, in recent years, peptide treatment has been found to be beneficial for a

wide variety of ailments. In the peptide treatment of non-viral diseases or conditions,
for example, it is believed that the peptides can be used to provide correct
information to cells for mammal cellular function, and thus better prepare the cells for
the treatment of the conditions and diseases. In viral treatments, the peptides can
act as protectors to the cellular surface by preventing the viral ligand proteins from
attaching to a receptor on a cellular surface.
[0065] The product can be administered as an oral spray or held in the mouth for
no less than 30 seconds to allow sorption through the mucous membrane of the
mouth. (For adults, with non-viral infections the composition is typically administered
twice a day wherein each administration consists of five successive 1 mL sprays; for
children, the composition is preferably administered twice a day wherein each
administration consists of two or three 1 mL sprays; for infants, the composition
preferably is administered twice a day wherein each administration consists of one 1
mL spray. For adults, with viral infections the composition is typically administered
every four hours wherein each administration consists of five successive 1 mL
sprays; for children, the composition is preferably administered every four hours
wherein each administration consists of two or three 1 mL sprays; for infants, the
composition preferably is administered every four hours wherein each administration
consists of one 1 mL spray.) Other administration routes include, without limitation,
injection, topical application, intraocular application, nebulization or atomization, and
the like. For example, a topical application of the composition might be indicated
when treating a burn or wound.
[0066] The following examples represent potential uses for peptides isolated
using the above process for treating a number of different medical conditions. It
should be noted that the isolated peptides can be used for numerous other
treatments and the examples listed are by no means exhaustive.
Examples
Example 1: Allergies
Typical dose: three sprays (5ml).
Typical administrations per day: 2
Typical interval of initial response: 1-3 days

Typical interval to benefit plateau: 3-7 days
Typical response: 2-3 days
No reduction or elimination of other therapeutics until justified by condition of patient.
Example 2: Arthritis
Typical dose: three sprays (5ml).
Typical administrations per day: 2
Typical interval of initial response: 7-21 days
Typical interval to benefit plateau: 42-56 days
Typical response: 2-3 days
No reduction or elimination of other therapeutics until justified by condition of patient.
Example 3: Benign Prostatic Hyperplasia (inflammatory
Aspect)
Typical dose: three sprays (5ml).
Typical administrations per day: 2
Typical interval of initial response: 7-14 days
Typical interval to benefit plateau: 14-28 days
Typical response: 2-3 days
No reduction or elimination of other therapeutics until justified by condition of patient.
Example 4: Cancer (Adjunctive use only)
Typical dose: three sprays (5ml).
Typical administrations per day: 2
Typical interval of initial response: 7-14 days
Typical interval to benefit plateau: n/a
Typical response: 2-3 days
No reduction or elimination of other therapeutics until justified by condition of patient.
Example 5: Celiac Sprue
Typical dose: three sprays (5ml).
Typical administrations per day: 2
Typical interval of initial response: 1 -3 days
Typical interval to benefit plateau: 3-7 days
Typical response: 2-3 days
No reduction or elimination of other therapeutics until justified by condition of patient.
Example 6: Crohn's Disease
Typical dose: three sprays (5ml).
Typical administrations per day: 2
Typical interval of initial response: 7-14 days
Typical interval to benefit plateau: 42-56 days
Typical response: 2-4 days
No reduction or elimination of other therapeutics until justified by condition of patient.
Example 7: Diabetes Type II
Typical dose: three sprays (5ml).
Typical administrations per day: 2

Typical interval of initial response: 1-7 days
Typical interval to benefit plateau: 14-28 days
Typical response: 2-3 days
No reduction or elimination of other therapeutics until justified by condition of patient.
Example 8: Hypertension
Typical dose: three sprays (5ml).
Typical administrations per day: 2
Typical interval of initial response: 7 days
Typical interval to benefit plateau: 28-56 days
Typical response: 2-3 days
No reduction or elimination of other therapeutics until justified by condition of patient.
Example 9: Lupus (Discoid and Systemic)
Typical dose: three sprays (5ml).
Typical administrations per day: 2
Typical interval of initial response: 7-14 days
Typical interval to benefit plateau: 28-58 days
Typical response: 2-3 days
No reduction or elimination of other therapeutics until justified by condition of patient.
Example 10: Multiple Sclerosis
Typical dose: three sprays (5ml).
Typical administrations per day: 2
Typical interval of initial response: 7-28 days
Typical interval to benefit plateau: 28-180 days
Typical response: 2-3 days
No reduction or elimination of other therapeutics until justified by condition of patient.
Example 11: Perthes Disease(Active)
Typical dose: three sprays (5ml).
Typical administrations per day: 2
Typical interval of initial response: 1-2 days
Typical interval to benefit plateau: 7-28 days
Typical response: 3-4 days
No reduction or elimination of other therapeutics until justified by condition of patient.
Example 12: Premenstrual Syndrome and Endometriosis
Typical dose: three sprays (5ml).
Typical administrations per day: 2
Typical interval of initial response: 1-2 days
Typical interval to benefit plateau: 14 days
Typical response: 2-4 days
No reduction or elimination of other therapeutics until justified by condition of patient.
Example 13: Prion Disease (Kuru and Creutzfeld-Jakob
Syndrome)
Typical dose: three sprays (5ml).

Typical administrations per day: 2
Typical interval of initial response: 12-28 days
Typical interval to benefit plateau: 28-56 days
Typical response: 2-3 days
No reduction or elimination of other therapeutics until justified by condition of patient.
Example 14: Psoriasis
Typical dose: three sprays (5ml).
Typical administrations per day: 2
Typical interval of initial response: 3-7 days
Typical interval to benefit plateau: 28-56 days
Typical response: 2-3 days
No reduction or elimination of other therapeutics until justified by condition of patient.
Example 15: Sjogren's Syndrome
Typical dose: three sprays (5ml).
Typical administrations per day: 2
Typical interval of initial response: 7-14 days
Typical interval to benefit plateau: 14-28 days
Typical response: 3-4 days
No reduction or elimination of other therapeutics until justified by condition of patient.
Example 16: Spinal Muscular Atrophy
Typical dose: three sprays (5ml).
Typical administrations per day: 2
Typical interval of initial response: 7-14 days
Typical interval to benefit plateau: 180 days
Typical response: 2-3 days
Increased consumption of water should be encouraged in adult patients with this
disease
Example 17: Thrombocytopenia (Ideopathic and
Autoimmune)
Typical dose: three sprays (5ml).
Typical administrations per day: 2
Typical interval of initial response: 1-2 days
Typical interval to benefit plateau: 4-10 days
Typical response: 3-4 days
No reduction or elimination of other therapeutics until justified by condition of patient.
Example 18: Topical Applications (bums, wounds, infections,
insect bites, diaper rash and Herpetic Lesions)
Typical method of administration Spray or apply with a sterile pad to affected
area.
Typical administrations per day: three sprays (3-5ml)
Typical interval to initial response: 4-24 hours
Typical interval to benefit plateau: Variable with condition
Typical response: 3-4 days

Typical observations include accelerated healing and reduced tendency to scar.
Example 19: Acute Viral Infections
Typical dose: three sprays (2ml).
Typical administrations per day: 6 (every four hours)
Typical interval of initial response: 4-24 hours
Typical interval to benefit plateau: 2-7 days
Typical response: 3-4 days
No reduction or elimination of other therapeutics until justified by condition of patient.
Example 20: Chronic Viral Infections (including HIV and
SARS)
Typical dose: three sprays (2ml).
Typical administrations per day: 6 (every four hours)
Typical interval of initial response: 4-24 hours
Typical interval to benefit plateau: 2-7 days
Typical response: 3-4 days
No reduction or elimination of other therapeutics until justified by condition of patient.
[0067] While the preferred embodiments of the invention have been illustrated
and described, it will be clear that the invention is not so limited. Numerous
modifications, changes, variations, substitutions and equivalents will occur to those
skilled in the art without departing from the spirit and scope of the present invention
as described in the claims.

WE CLAIM:
1. A method for separating the proline-rich polypeptides essentially free of
peptides and proteins of about 10,000 Daltons or greater from the globular
proteins having a molecular weight of between 10,000 and 500,000
Daltons in bovine colostrum, the method comprising the steps of: (a)
subjecting at least a portion of a bovine colostrum sample to at least one
separation procedure comprising an ion exchange separation to yield
separated proline-rich polypeptides essentially free of peptides and
proteins of about 10,000 Daltons or greater and separated globular
proteins having a molecular weight of between 10,000 and 500,000
Daltons, wherein no more than about 10% of the separated proline-rich
polypeptides are denatured; and (b) collecting the separated proline-rich
polypeptides; wherein at least 90% of the separated proline-rich
polypeptides have a molecular weight of no more than about 2,500
Daltons.
2. The method as claimed in claim 1, wherein the step (a) is performed at a
pressure of 30 psi or less.
3. The method as claimed in claim 1, wherein the step (a) is performed at a
pressure of 10 psi or less.

4. The method as claimed in claim 1, wherein the at least one separation
procedure further comprises a size exclusion separation.
5. The method as claimed in claim 1, wherein the step (a) comprises
changing the pH of the at least a portion of the colostrum.


A method for separating the proline-rich polypeptides essentially free of peptides
and proteins of about 10,000 Daltons or greater from the globular proteins having
a molecular weight of between 10,000 and 500,000 Daltons in bovine colostrum,
the method comprising the steps of: (a) subjecting at least a portion of a bovine
colostrum sample to at least one separation procedure comprising an ion
exchange separation to yield separated proline-rich polypeptides essentially free
of peptides and proteins of about 10,000 Daltons or greater and separated
globular proteins having a molecular weight of between 10,000 and 500,000
Daltons, wherein no more than about 10% of the separated proline-rich
polypeptides are denatured; and (b) collecting the separated proline-rich
polypeptides; wherein at least 90% of the separated proline-rich polypeptides
have a molecular weight of no more than about 2,500 Daltons.

Documents:

00942-kolnp-2006-abstract.pdf

00942-kolnp-2006-claims-1.1.pdf

00942-kolnp-2006-claims.pdf

00942-kolnp-2006-correspondence others-1.1.pdf

00942-kolnp-2006-correspondence-1.2.pdf

00942-kolnp-2006-cprrespondence other.pdf

00942-kolnp-2006-description (complete).pdf

00942-kolnp-2006-drawings.pdf

00942-kolnp-2006-form-1-1.1.pdf

00942-kolnp-2006-form-1.pdf

00942-kolnp-2006-form-13.pdf

00942-kolnp-2006-form-2.pdf

00942-kolnp-2006-form-26.pdf

00942-kolnp-2006-form-3.pdf

00942-kolnp-2006-form-5.pdf

00942-kolnp-2006-fprm-18.pdf

00942-kolnp-2006-international publication.pdf

00942-kolnp-2006-international search authority report.pdf

00942-kolnp-2006-pct form-1.1.pdf

00942-kolnp-2006-pct form.pdf

00942-kolnp-2006-pct others.pdf

00942-kolnp-2006-priority document.pdf

942-KOLNP-2006-ABSTRACT 1.1.pdf

942-KOLNP-2006-ABSTRACT 1.2.pdf

942-KOLNP-2006-CANCELLED PAGES.pdf

942-KOLNP-2006-CLAIMS 1.1.pdf

942-KOLNP-2006-CLAIMS 1.2.pdf

942-KOLNP-2006-CORRESPONDENCE-1.2.pdf

942-kolnp-2006-correspondence.pdf

942-KOLNP-2006-DESCRIPTION (COMPLETE) 1.1.pdf

942-KOLNP-2006-DESCRIPTION (COMPLETE) 1.2.pdf

942-KOLNP-2006-DRAWINGS 1.1.pdf

942-kolnp-2006-examination report.pdf

942-KOLNP-2006-FORM 1 1.2.pdf

942-KOLNP-2006-FORM 1.1.1.pdf

942-kolnp-2006-form 1.pdf

942-kolnp-2006-form 13.pdf

942-kolnp-2006-form 18.pdf

942-KOLNP-2006-FORM 2 1.2.pdf

942-KOLNP-2006-FORM 2.1.1.pdf

942-kolnp-2006-form 26.pdf

942-KOLNP-2006-FORM 3 1.2.pdf

942-KOLNP-2006-FORM 3.1.1.pdf

942-kolnp-2006-form 3.pdf

942-KOLNP-2006-FORM 5.1.1.pdf

942-kolnp-2006-form 5.pdf

942-KOLNP-2006-FORM-27-1.pdf

942-KOLNP-2006-FORM-27.pdf

942-kolnp-2006-granted-abstract.pdf

942-kolnp-2006-granted-claims.pdf

942-kolnp-2006-granted-description (complete).pdf

942-kolnp-2006-granted-drawings.pdf

942-kolnp-2006-granted-form 1.pdf

942-kolnp-2006-granted-form 2.pdf

942-kolnp-2006-granted-specification.pdf

942-KOLNP-2006-OTHERS 1.1.pdf

942-kolnp-2006-others-1.2.pdf

942-KOLNP-2006-OTHERS.pdf

942-KOLNP-2006-PA 1.1.pdf

942-KOLNP-2006-PA.pdf

942-KOLNP-2006-PETITION UNDER RULE 137.pdf

942-KOLNP-2006-REPLY TO EXAMINATION REPORT 1.1.pdf

942-kolnp-2006-reply to examination report-1.2.pdf

942-KOLNP-2006-REPLY TO EXAMINATION REPORT.pdf

abstract-00942-kolnp-2006.jpg


Patent Number 247492
Indian Patent Application Number 942/KOLNP/2006
PG Journal Number 15/2011
Publication Date 15-Apr-2011
Grant Date 12-Apr-2011
Date of Filing 17-Apr-2006
Name of Patentee ADVANCED PROTEIN SYSTEMS
Applicant Address 601 SOUTH 54TH AVENUE, SUITE 101, PHOENIX, AZ 85043
Inventors:
# Inventor's Name Inventor's Address
1 KEECH ANDREW 8300 EAST VIA DE VENTURA, #1032, SCOTTSDALE, AZ
2 JIMENEZ-FLORES RAFAEL 4444 SANFLOWER WAY, SAN LUIS OBISPO, CA 93401
PCT International Classification Number A23J
PCT International Application Number PCT/US2004/036566
PCT International Filing date 2004-11-03
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 60/516,674 2003-11-03 U.S.A.