Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Monoclonal antibody of anti human immune deficiency virus I type 24 protein and application thereof

A technology of immunodeficiency virus and monoclonal antibody, which is applied in the direction of antiviral immunoglobulin, measuring devices, instruments, etc., can solve the problems of no public reports and increase of non-specific reactions, and achieve high sensitivity, strong specificity, and preparation The effect of simple method

Inactive Publication Date: 2005-10-26
KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
View PDF1 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

U.S. Patent Publication No. US 004,886,742, published on December 12, 1989, the name of the invention is Enzyme immunoassay for detecting HIV antigens in human sera, and the application discloses a method for detecting human immunodeficiency virus p24 protein, which has the disadvantage The point is to use human positive serum as the detection antibody, which increases the possibility of non-specific reaction
[0007] Through document retrieval, do not see the public report of identical document of the present invention

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Monoclonal antibody of anti human immune deficiency virus I type 24 protein and application thereof
  • Monoclonal antibody of anti human immune deficiency virus I type 24 protein and application thereof
  • Monoclonal antibody of anti human immune deficiency virus I type 24 protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1, the preparation of monoclonal antibody

[0037] (1) Immunized mice

[0038] The immunogen used to prepare the monoclonal antibody is the purified subtype B HIV-1p24 genetically engineered recombinant protein. The full-length protein of subtype B HIV-1p24 was expressed in E.coli expression system expressing HIV-1B subtype p24 protein in large quantities, and the GST-p24 fusion protein was expressed in large quantities. After the bacteria were lysed with lysozyme, the Parmacia Sepharose-4B GST affinity chromatography column was used. GST fusion protein was adsorbed, digested with PreScission protease at 4°C overnight, and eluted with elution buffer to obtain electrophoretic pure HIV-1B subtype recombinant p24 protein.

[0039] BALB / c mice were immunized with subtype B HIV-1p24 recombinant protein, and the specific immunization scheme was as follows:

[0040] HIV-1p24 antigen plus complete Freund's adjuvant for initial immunization, 50ug / body

[0041] ...

Embodiment 2

[0060] Embodiment 2, identification of monoclonal antibody

[0061] fusion cell chromosome count

[0062] The confluent cells cultured into a single layer were cultured with colchicine (0.1 μg / ml) for 36-48 hours, treated with 0.075 M KCI for hypotonicity, fixed with glacial acetic acid and methanol, prepared into slices and stained with Giemsa. The chromosomes of 20 effective cells were counted in each of the three hybridomas. The average number of chromosomes in hybridoma p3JB9 was 89.7±5.5, the average number of chromosomes in p5F1 was 91.0±1.5 and the average number in p6F4 was 83.2±4.6.

[0063] Karyotype analysis of three hybridoma cells figure 1 shown.

[0064] Identification of monoclonal antibody Ig class / subclass

[0065] With plating solution (Na 2 CO 3 1.59g / l, NaHCO 3 2.93g / l, pH 9.6) Dilute the p24 antigen to 1μg / ml, 100μl / well, overnight at 4°C; wash the plate with washing solution, seal the plate with blocking solution (5% milk powder, PBS, pH 7.4), and...

Embodiment 3

[0079] Example 3, mass production of monoclonal antibodies

[0080] First intraperitoneally inject 0.5ml Pristane (pristane) to BALB / c mice, and intraperitoneally inject O.5×10 6 Ascites can be produced 7-10 days after the inoculation of the cells, the mice were sacrificed, the ascites was extracted with a syringe, the supernatant was obtained by centrifugation, and stored at -20°C. The content of monoclonal antibody in ascites can reach 5-20mg / ml.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The present invention to a monoclonal antibody for resisting human immune deficiency virus I type p24 protein and its application, belonging to the field of biological technology. The immunogen of monoclonal antibody is HIV-IB substype p24 gene engineering recombinant protein. Said monoclonal antibody can be combined with other monoclonal antibody or polyclonal antibody or can be used as label of radioactive isolope, enzyme, fluorescin compound, chemical luminescence comound or colloidal metal ion, can be used for preparing various qualitative or quantitative detection liquors and reagent of p24 antigen in cultured supernatant fluid or cell or tissue.

Description

Technical field: [0001] The present invention relates to a kind of anti-human immunodeficiency virus type I (HIV-1) p24 protein monoclonal antibody and application thereof, more specifically utilizes genetic engineering to prepare recombinant HIV-1p24 protein and its monoclonal antibody, and is used for HIV - Detection of 1p24 antigen. Background technique: [0002] AIDS (Acquired Immunodeficiency Syndrome, AIDS) is widely prevalent in the world, and Asia is one of the regions where HIV (Human Immunodeficiency Virus) infection rises most rapidly and is most prevalent after North America and Africa. my country's AIDS also showed a trend of accelerated prevalence. Experts estimate that the actual number of HIV-infected people in my country has exceeded 1 million. By the end of 2003, the number of existing HIV-infected people was estimated to be 840,000. As of the end of September 2004, there were 89,067 reported cases of HIV infection, including 20,786 cases of AIDS. [000...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/08
Inventor 郑永唐刘广杰杨柳萌
Owner KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com