Title of Invention | A DEVICE FOR DETECTION OF ANTIBODIES TO HIV AND p24 ANTIGEN OF HIV 1 IN HUMAN SERUM OR PLASMA |
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Abstract | The present invention HIV Ag & Ab Comb is a test device for the early detection of p24 antigen and HIV1 and HIV 2 antibodies in human serum or plasma or whole blood. It is a visual, rapid, sensitive and accurate test kit for analyzing the presence of HIV antibodies and he workability of the device can be tested simultaneously with the detection of the presence of p24 antigen and antibodies to HIV. |
Full Text | The present invention relates to the device for the determination of the HIV in human serum / plasma/ whole blood. More specifically the present invention relates to the process for making the HIV Comb for the early detection of HIV1 and HIV 2 antibodies and HIV p 24 antigen in human serum or plasma or whole blood. The present invention relates to detection kit for the early detection of the presence of HIV 1 and HIV 2 antibodies and p24 antigen in human serum or plasma or whole blood. HIV Comb+ Ag is a visual and rapid, sensitive and accurate test device for analyzing the presence of HIV 1 and HIV 2 antibodies and HIV p 24 antigen in human serum or plasma or whole blood. It is a test device wherein the presence of antibodies to HIV1, HIV2 and p24 antigen and workability of the test device is determined separately and simultaneously. Moreover the early detection is extremely important for all the patients regardless of lifestyle in reducing the further transmission. Back ground HIV stands for Human Immunodeficiency Virus. HIV is the virus that causes AIDS. Its transfusion is resulting from blood transfusion. AIDS disease, which is primarily caused by infection of individuals with a HIV retrovirus, is now the most devastating disease in the whole world, since the number of individuals which are, to date, infected with HIV viruses is estimated to about 40 millions of individuals. During the year 2001, 5 millions of individuals were infected with HIV while 3 millions of individuals have deceased in the same time. Since the discovery of the main AIDS causative agent in 1983, namely the HIV virus, extensive efforts have been made in order to understand the mechanism of action of this virus and to develop accurate methods for 2 (i) reproducibly diagnosing the infection, as well as (ii) carrying out a prognosis of the progression of the disease in a given patient. There are two kinds of viruses, HIV 1 and HIV 2 which causes the infection. These two viruses are retroviruses group and are slow viruses. This knowledge has laid a foundation for the development of a new assay based on the Recombinant DNA technology leading to the differential detection of antibodies to HIV-1 and HIV-2 in human serum or plasma. Prior Arts US Patent No: 6870045: describes the kit for detecting HIV-2. It describes the compositions and method for synthesizing and detecting HIV-2 specific amplicons. Particularly described are oligonucleotides that are useful as hybridization probes, and amplification primers that facilitate detection of very low levels of HIV-2 nucleic acids. US Patent No. 20060257318 describes the field of diagnosis of the progression status of an infection of an individual with a virus belonging to the family of HIV as well as with the therapeutical treatment of this infection disease. US Patent No.20060234209: describes the microchip based assays to measure HIV associated analysts of interest in a sample from a subject infected with the HIV virus. Methods of present invention are optimal for use in monitoring HIV disease in resource poor settings. US Patent No: 7115363: The invention relates to a new class of retroviruses, designated by HIV-2, of which samples have been deposited to the ECACC under numbers 87.01.1001 and 87.01.1002 and to the NCIB under numbers 12.398 and 12.399. It relates to antigens capable to be obtained from this virus, particularly proteins p12, p16, p26 and gpl40. These various antigens can be used for the diagnosis of the disease, especially by contacting these antigens with a serum of a patient submitted to the diagnosis. It relates to immunogenic compositions containing more particularly the glycoprotein gpl40. Finally it concerns nucleotidic sequences, which can be used especially as hybridization probes, derived from the RNA of HIV-2. 3 US Patent No: 7083922: relates to oligonucleotides for use in amplifying and detecting HIV nucleic acid in a sample. Description HIV p24 antigen test device for the detection of the specific p24 antigen is a test device for the early and rapid detection of the HIV in human serum or plasma. Antigen can generally only be detected during the acute and during the symptomatic phase of AIDS. Antibodies to HIV can be detected throughout virtually the entire infection period. When the HIV virus enters into the body, the earlier detection of the disease is impossible because the virus enters in window phase and subsequently the immune system start producing antibodies. So the use of highly sensitive antibody assay is therefore an established approach in serodiagnosis of HIV infection. Progressive improvements in assay sensitivity have reduced that window phase. Further shortening of the period can be achieved by the incorporation of HIV antigen detection in such antibody tests enabling the detection of the infected individuals at the earliest possible moment. P24 detection dot detects the specific p24 antigen of HIV. So early detection of HIV is possible. The present invention relates to a test device for the earlier detection of p24 antigen and antibodies to HIV1 & 2. It comprises of a comb, which is made of polystyrene plastic sheet. The plastic was die-cut into "combs" having eight individual "teeth," each a separate test, spaced to fit into a standard 96-well micro test plate in which an assay is performed. It has three dots on the side of the each tooth, of which the upper dot is an inbuilt control dot, which determines the process done is perfectly right. The central dot is the dot for the determination of the p24 antigen and the lowest dot is for the determination of the HIV-1 or HIV -2 antibodies. These three antigen and antibody dots are immobilized as circular coats on the comb. The present invention relates to a rapid, visual, sensitive and qualitative in vitro diagnostic kit for the detection of the HIV 1 and HIV 2 antibodies and p24 antigens of HIV in human serum and plasma and whole blood comprising a test polystyrene comb, wash buffer solution, sample diluent and conjugates. 4 There are five formats by which the test can be performed. Interpretations of results for the five formats are same as described below. On performing the test, if only the control dot appears which is pink in colour, then the result is interpreted as negative and the procedure carried out for the detection are perfectly right. If both the control dot and HIV-1 and HIV-2 antibody detection dot appears in pink colour, then the result is that the patient's sample is reactive for HIV antibody. If the control dot and p24 antigen detection dot appear in pink colour, then the patient's sample is reactive for HIV p24 antigens. If the control dot, HIV p24 detection dot and the HIV antibody dot appear pink in colour, then the patient's sample is reactive for HIV antibody and p24 antigen. The control dot to test the workability of the device is coated with antihuman IgG for eg. rabbit, goat, donkey or chicken. HIV1 and HIV2 detection dot is coated with a mixture of glycoprotein gp41, C terminus of glycoprotein gpl20 and glycoprotein gp36 which is immobilized on the comb in the form of dot. p24 dot for the detection of p24 antigen is coated with specific monoclonal or polyclonal or F(ab1)2 p24 antibodies. The quantity used for the coating is 1 nanogram to 10 microgram for each protein. Sample The sample should be only human serum or plasma/ whole blood. Principle HIV-1 and HIV-2 antigens and p24 antibodies are immobilized on the polystyrene comb. When the patient's sample is added to the device, HIV antibodies and p24 antigens if present, bind to the coated antigens and p24 antibodies respectively. P24 antibody conjugate is added which binds to the p24 antibody- antigen complex and protein A conjugate binds to the Fc portion of HIV antibodies to give pinkish purple dots against white background. 5 In an embodiment of the present invention, the process for making the HIV Ag & Ab comb for analyzing the presence of HIV antibodies and p24 antigen in human serum or plasma is as follows: Preparation of HIV1 and HIV2 detection dot To 5 mM - 100 mM carbonate buffer solution of pH 8- 9.6, add 1 nano gram to 10 micro gram of each glycoprotein gp 41, C terminus of glycoprotein 120 and glycoprotein 36 and vortex the mixture to get a homogeneous solution. Coat 1 Μ1 — 10 μl of the above mixture to the HIV 1 and HIV 2 detection dot and allow it to dry at 37 ° C for 1 to 20 hrs and preferably 4 hrs. Preparation of Control Dot To 5 mM -100 mM carbonate buffer solution of pH 8- 9.6, add 1 nano gram to 10 micro gram antihuman IgG for eg. of chicken or goat and vortex the mixture to get a homogeneous solution. Coat 1 ΜL — 10 ul of the above mixture to the control dot and allow it to dry at 37 ° C for 1 to 20 hrs and preferably 4 hrs. Preparation of p24 detection dot To 5 mM - 100 mM carbonate buffer solution of pH 8- 9.6, add 1 nano gram to 10 micro gram specific monoclonal or polyclonal or F(ab )2 p24 antibodies and vortex the mixture to get a homogeneous solution. Coat 1 ul - 10 ul of the above mixture to the p24 antigen detection dot and allow it to dry at 37 ° C. In an embodiment of the present invention, preparation of sample diluent is as follows. To 10-100 mM Phosphate Buffer Saline of pH 7-8 , add detergent 0.01 % - 0.5% and dye. In an another embodiment the wash buffer solution is prepared by adding 50- 500 mM Phosphate Buffer Solution of pH 6.5 - 8.5 to the detergent of 0.005 % - 0.5 % . 6 Example; 1 Preparation of protein A conjugate 1. To an initial solution of 0.01 % - 0.05% (w/v) Gold Chloride (HAuCl4) that is at boiling temperature, sodium citrate solution (1-5%) is added . The solution is rapidly stirred and boiled until a pink purple colour develops. Protein A is added to the same. 2. Any non- immune animal serum (mouse / goat / rabbit /sheep) is added to the above solution. The concentration of serum could be lμg to 10μg /ml of gold colloid solution. This solution is mixed for 2 hr to over night. After this the solution is centrifuged and the supernatant is discarded. Dissolve the pellet in buffer containing stabilizers such as BSA, Gelatin to obtain the Protein A conjugate. Preparation of P24 conjugate. 1. To an initial solution of 0.01 % - 0.05% (w/v) Gold Chloride (HAuCl4) that is at a boiling temperature, sodium citrate solution (1-5%) is added . The solution is rapidly stirred and boiled until a pink purple colour develops. P24 antibody monoclonal / polyclonal is added to the same. Adjust the pH of the gold Colloid to 0.3 units higher than pI of p24 antibody monoclonal or poly clonal. 2. Any non- immune animal serum (mouse / goat / rabbit /sheep) is added to the above solution. The concentration of serum be 1μg tol0μg /ml of gold colloid solution. This solution is mixed from 2 hrs to overnight. After this the solution is centrifuged and the supernatant is discarded. Dissolve the pellet in buffer containing stabilizers such as BSA, Gelatin to obtain the P24 conjugate. P 24 conjugate is in lyophilized or liquid state. 7 Example 2; Preparation of HIV 1 and HIV 2 conjugate. 1. To an initial solution of 0.01 % - 0.05% (w/v) Gold Chloride (HAucl4) that is at a boiling temperature, sodium citrate solution (1-5%) is added . The solution is rapidly stirred and boiled until a pink purple colour develops. Adjust the pH of the Gold Chloride equal to 0.3 units higher than pi of Ag. Recombinant/ synthetic peptides for HIV 1 and HIV2 is added to the same and mixed for 30 minutes to 1 hour . IgG from the species corresponding to the control dot is added and mixed for one hour. 2. Any non- immune animal serum (mouse or goat or rabbit sheep) is added to the above solution. The concentration of serum could be lug tol0μg /ml of gold colloid solution. This solution is mixed for 2 hrs to overnight. After this the solution is centrifuged and the supernatant is discarded. Dissolve the pellet in buffer containing stabilizers such as BSA, Gelatin to obtain the HIV 1 & 2 conjugate. HIV 1 and HIV2 Conjugate is in lyophilized or liquid state. Example 3: Monoclonal or Polyclonal antihuman IgG gold conjugate 1. To an initial solution of 0.01 % - 0.05% (w/v) gold choride ( HAuC14) that is at boiling temperature, sodium citrate solution (1-5%) is added. The solution is rapidly stirred and boiled until a pink purple colour develops. Adjust the pH of the gold colloid solution to 0.3 units higher than the PI of antibody. Monoclonal or Polyclonal antihuman IgG is added to the same. 1. Any non - immune animal serum ( mouse/ goat/ rabbit/ sheep) is added to the above solution. The concentration of the serum could be 1 ul to 10|al/ml of gold colloid solution. This solution is mixed for 2 hr to overnight. After this the 8 solution is centrifuged and supernatant is discarded. Dissolve the pellet in buffer containing stabilizers such as BSA, Gelatin to obtain the antihuman IgG Gold conjugate. In another embodiment of the present invention, test procedure for analyzing the presence of HIV antibodies and p24 antigen in human serum and plasma is as follows: 1. Taking 150 μl of sample in the microcuvettes and add 150μl of sample diluent to it. 2. Mixing the sample with the sample diluent repeatedly. Make sure that no bubbles are formed while mixing the sample. 3. Cutting the pouch and take out the necessary no of combs and place it into the microcuvettes-containing sample and incubate for 10 minutes at room temperature. During incubation withdrawing and inserting the comb in the microcuvettes for 15 seconds. 4. Removing the comb from the sample and blot the surface of the comb. 5. Washing the comb in the wash buffer solution for 10 times carefully by moving the comb forward and back ward in the wash solution. 6. Placing the comb in the microcuvettes containing p24 conjugate and incubating it for 10 minutes. Withdrawing and inserting the comb in the solution for 15 seconds 7. Washing the comb in the wash buffer solution. 8. Immersing the comb in to the microcuvettes containing protein A conjugate and incubating for 10 minutes in room temperature. Withdrawing and inserting the comb in to the microcuvettes containing protein A conjugate for 15 seconds. 9. Taking the comb out and placing the comb on a clean surface, reactive side up and allowing it to dry. 10. Reading the result only after the comb is completely dry. 9 Example 4 Preparation of Streptavidin Gold conjugate. 1. To an initial solution of 0.01 % - 0.05% (w/v) Gold Chloride (HAuCl4) that is at a boiling temperature, sodium citrate solution (1-5%) is added . The solution is rapidly stirred and boiled until a pink purple colour develops. Streptavidin is added to the same. 2. Any non- immune animal serum (mouse / goat / rabbit /sheep) is added to the above solution. The concentration of serum could be lμg tol0μg /ml of gold colloid solution. This solution is mixed for 2 hrs to overnight. After this the solution is centrifuged and the supernatant is discarded. Dissolve the pellet in buffer containing stabilizers such as BSA, Gelatin to obtain the Streptavidin gold conjugate. Biotinvlated solution of HIV 1. HIV2 and p24 Biotinylated solution of HIV1, HFV2 antigen and p24 antibody can be prepared using commercially available biotinylant kit. Test procedure 1. Taking 150 ul of sample in the microcuvettes and add 150μl of sample diluent to it. 2. Mixing the sample with the sample diluent repeatedly. Make sure that no bubbles are formed while mixing the sample. 3. Cutting the pouch and taking out the necessary no of combs and place it into the microcuvettes-containing sample and incubating for 10 minutes at room 10 temperature. During incubation withdrawing and inserting the comb in the microcuvettes for 15 seconds. 4. Removing the comb from the sample and blot the surface of the comb. 5. Washing the comb in the wash buffer solution for 10 times carefully by moving the comb forward and back ward in the wash solution. 6. Immersing the comb in to the microcuvettes containing biotinylated HIV1 HIV2 antigen and p24 antibody and incubating for 10 minutes in room temperature. Withdrawing and inserting the comb in to the microcuvettes for 15 seconds. 7. Washing the comb in the wash buffer solution for 10 minutes carefully by moving the comb forward and backward in the wash solution. 8. Placing the comb in the microcuvettes containing Streptavidin Gold Conjugate and incubating it for 10 minutes. Withdrawing and inserting the comb in the solution for 15 seconds. 9. Taking the comb out and placing the comb on a clean surface, reactive side up and allowing it to dry. 10. Reading the result only after the comb is completely dry. According to the invention, a device for an early detection of the p24 antigen and antibodies to HIV in the human beings, comprises of a polystyrene HIV comb having 8 teeth or comb, provided with three dots on each teeth or comb, the upper dot is an inbuilt control dot, middle dot for the detection of p24 antigen and the lower dot for the detection of HIV antibodies, wherein the dots for the detection of HIV is coated with homogeneous solution of HIV antigen and control dot is coated with antihuman IgG and the p24 detection dot is coated with monoclonal or polyclonal or (Fab!)2 p24 antibodies. According to the present invention, A process for making the HIV Ag & Ab Comb device for analyzing the presence of HIV antibodies and p24 antigen in human serum or plasma comprises the following steps: a. adding 5 mM - 100 mM carbonate buffer solution of pH 8- 9.6, 1 nano gram to 10 micro To gram glycoprotein gp 41, C terminus of glycoprotein 120 and glycoprotein 36, vortexing the mixture to get a homogeneous solution, coating 1 μl— 10 μl of the above mixture to the HIV 1 and HIV 11 2 detection dot and allow it to dry at 37°C for 1 hour to 20 hours and preferably 4 hrs. b. to 5 mM - 100 raM carbonate buffer solution of pH 8- 9.6, adding 1 nano gram to 10 micro gram antihuman IgG for eg. of chicken or goat vortexing the mixture to get a homogeneous solution, Coating 1 μl - 10 ul of the above mixture to the control dot and allowing it to dry at 37 ° C for 1 hr to 20 hrs and preferably 4 hrs. c. To 5 mM -100 mM carbonate buffer solution of pH 8- 9.6, adding 1 nano gram to 10 micro gram specific monoclonal or polyclonal p24 antibodies vortexing the mixture to get a homogeneous solution, Coating 1μl— 10 ul of the above mixture to the p24 antigen detection dot and allowing it to dry at 37 ° C for 1 hr to 20 hrs and preferably 4 hrs . According to the invention, a device as claimed in claim 1 wherein the device is adapted to analyze as follows:- a. taking 150 μl of sample in the microcuvettes and adding 150ul of sample diluent to it. b. mixing the sample with the sample diluent repeatedly avoiding formation of bubbles while mixing the sample. c. cutting the pouch and taking out the necessary no of combs and placing it into the microcuvettes-containing sample and incubate for 10 minutes at room temperature, d. during incubation withdrawing and inserting the comb in the microcuvettes for 15 seconds. e. removing the comb from the sample and blot the surface of the comb. f. washing the comb in the wash buffer solution for 10 times carefully by moving the comb forward and back ward in the wash solution. g. placing the comb in the microcuvettes containing p24 conjugate and incubate it for 10 minutes at room temperature. h. withdrawing and inserting the comb in the wash buffer solution for 15 seconds 12 i. washing the comb in the wash buffer solution for 10 times carefully by moving the comb forward and back ward in the wash solution. j. immersing the comb in to the microcuvettes containing protein A conjugate and incubate for 10 minutes in room temperature, withdrawing and inserting the comb in to the microcuvettes containing protein A conjugate for 15 seconds. k. Placing the comb on a clean surface, reactive side up and allow it to dry. 1. Reading the result only after the comb is completely dry. The invention will be described with reference to the following accompanying drawings wherein Figure 1 shows the fourth generation of the HIV Ag/ Ab Comb. This figure clearly shows the difference in the various devices developed from time to time. In the first generation, the antibodies were detected only after 6 weeks. In the second generation, the antibodies were detected before 6 weeks but after 4 weeks. In the third generation, the antibodies were detected after three weeks but before 4 weeks. In the fourth generation, the antigens were detected before three weeks but after 2 weeks. So this is an important invention in the history of HIV. Figure 2 shows the polystyrene comb. Figure 3 shows the workability of the device is perfectly right. Figure 4 shows the presence of HIV antibodies in the human serum or plasma and the device is working properly. Figure 5 shows the presence of p24 antigen in the human serum or plasma and the workability of the device. Figure 6 shows the presence of HIV antibodies and p24 antigen in the human serum or plasma and the workability of the device. Figure 7 shows the absence of HIV antigen and antibodies. 13 I Claim:- 1. A device for an early detection of the p24 antigen and antibodies to HIV in the human beings, comprises of a polystyrene HIV comb having 8 teeth or comb, provided with three dots on each teeth or comb, the upper dot is an inbuilt control dot, middle dot for the detection of p24 antigen and the lower dot for the detection of HIV antibodies, wherein the dots for the detection of HIV is coated with homogeneous solution of HIV antigen and control dot is coated with antihuman IgG and the p24 detection dot is coated with monoclonal or polyclonal or (Fab')2 p24 antibodies. 2. A device as claimed in claim 1 wherein the dot for the detection of HIV 1 and HIV 2 antibodies are coated with glycoprotein gp41, C terminus of gp 120 and gp 36 glycoprotein. 3. A device as claimed in claim 1 where in the antigens used for the coating are gp 41 , C terminus of gp 120 and gp 36. 4. A device as claimed in claim 1 wherein the control dot is coated with antihuman IgG for example of rabbit, goat, donkey or chicken. 5. A device as claimed in claim 1 wherein the p24 detection dot is coated with specific monoclonal or polyclonal or (Fab1)2 p 24 antibodies. 6. A device as claimed in claim 1 wherein 1 nano gram to 10 micro gram p24 antibodies are used for coating the p24 detection dot. 7. A process for making the HIV Ag & Ab Comb device for analyzing the presence of HIV antibodies and p24 antigen in human serum or plasma comprises the following steps: a. adding 5 mM - 100 mM carbonate buffer solution of pH 8- 9.6, 1 nano gram to 10 micro To gram glycoprotein gp 41, C terminus of glycoprotein 120 and glycoprotein 36, vortexing the mixture to get a homogeneous solution, coating 1 ul - 10 ΜL of the above mixture to the HIV 1 and HIV 2 detection dot and allow it to dry at 37°C for 1 hour to 20 hours and preferably 4 hrs. b. to 5 mM - 100 mM carbonate buffer solution of pH 8- 9.6, adding 1 nano gram to 10 micro gram antihuman IgG for eg. of chicken or goat 14 vortexing the mixture to get a homogeneous solution, Coating 1 μ1 — 10 μl of the above mixture to the control dot and allowing it to dry at 37 ° C for 1 hr to 20 hrs and preferably 4 hrs. c. To 5 mM -100 mM carbonate buffer solution of pH 8- 9.6, adding 1 nano gram to 10 micro gram specific monoclonal or polyclonal p24 antibodies vortexing the mixture to get a homogeneous solution, Coating 1 μ1 - 10 μ1 of the above mixture to the p24 antigen detection dot and allowing it to dry at 37 ° C for 1 hr to 20 hrs and preferably 4 hrs . 8. A device as claimed in claim 1 wherein the device is adapted to analyze as follows:- a. taking 150 μl of sample in the microcuvettes and adding 150μl of sample diluent to it. b. mixing the sample with the sample diluent repeatedly avoiding formation of bubbles while mixing the sample. c. cutting the pouch and taking out the necessary no of combs and placing it into the microcuvettes-containing sample and incubate for 10 minutes at room temperature, d. during incubation withdrawing and inserting the comb in the microcuvettes for 15 seconds. e. removing the comb from the sample and blot the surface of the comb. f. washing the comb in the wash buffer solution for 10 times carefully by moving the comb forward and back ward in the wash solution. g. placing the comb in the microcuvettes containing p24 conjugate and incubate it for 10 minutes at room temperature. h. withdrawing and inserting the comb in the wash buffer solution for 15 seconds i. washing the comb in the wash buffer solution for 10 times carefully by moving the comb forward and back ward in the wash solution . j. immersing the comb in to the microcuvettes containing protein A conjugate and incubate for 10 minutes in room temperature, withdrawing 15 and inserting the comb in to the microcuvettes containing protein A conjugate for 15 seconds. k. Placing the comb on a clean surface, reactive side up and allow it to dry. 1. Reading the result only after the comb is completely dry. 9. A device for an early detection of the p24 antigen and antibodies to HIV in the human beings substantially as hereinbefore described with reference to the drawings. 10. A process for making the HIV Ag & Ab Comb device for analyzing the presence of HIV antibodies and p24 antigen in human serum or plasma substantially as hereinbefore described with reference to the drawings. 11. A device to analyze the p24 antigen & antibidoes to HIV substantially as hereinbefore described with reference to the drawings. Dated this 4th day of May, 2007. 16 The present invention HIV Ag & Ab Comb is a test device for the early detection of p24 antigen and HIV1 and HIV 2 antibodies in human serum or plasma or whole blood. It is a visual, rapid, sensitive and accurate test kit for analyzing the presence of HIV antibodies and p24 antigen in human serum or plasma or whole blood. In this test device, the workability of the device can be tested simultaneously with the detection of the presence of p24 antigen and antibodies to HIV. |
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Patent Number | 237854 | ||||||||
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Indian Patent Application Number | 693/KOL/2007 | ||||||||
PG Journal Number | 04/2010 | ||||||||
Publication Date | 22-Jan-2010 | ||||||||
Grant Date | 11-Jan-2010 | ||||||||
Date of Filing | 07-May-2007 | ||||||||
Name of Patentee | MAHAJAN; LALIT | ||||||||
Applicant Address | 1-D, MANHAR MAHAL, 4, BAKUL BAGAN ROA BEHIND LANSDOWNE MKT., KOLKATA - 700 025 | ||||||||
Inventors:
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PCT International Classification Number | C07K16/10 | ||||||||
PCT International Application Number | N/A | ||||||||
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