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A broad-spectrum neutralizing antibody against HIV

An antibody and carrier technology, applied in the field of cellular immunology and genetic engineering, can solve the problems of virus mutation and poor immunogenicity, and achieve high success rate and good application prospects

Active Publication Date: 2021-06-18
SUZHOU HEALTH COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

First, if the induced antibody wants to have a broad spectrum and be able to neutralize a variety of HIV strains, the antibody must be able to directly recognize conserved epitope sites, and these sites often exist in the envelope protein of HIV and are covered by polysaccharides. Region coverage, resulting in close confinement of these buried conserved sites, resulting in poor immunogenicity
Second, the virus can mutate quickly in the body, thus permanently escaping the detection of the immune system

Method used

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  • A broad-spectrum neutralizing antibody against HIV
  • A broad-spectrum neutralizing antibody against HIV
  • A broad-spectrum neutralizing antibody against HIV

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1 Preparation of an anti-HIV antibody

[0068] 1, sorting of mature B cells

[0069] Buffer1: 500ml 1 × PBS + 2mL 0.5m EDTA + 25ml 10% BSA (including 2mm EDTA, 0.5% BSA)

[0070] (1) Collect people from blood of 200 μl of blood, 200 μl of BUFFER1;

[0071] (2) Add 10x volumes of Ack Lysing Buffer (Fisher / Biowhittaker), 10 min, 1500 rpm for 10min, 1500 rpm;

[0072] (3) Abandon the discard, add 10ml buffer 1 washing cells, 1500 rpm centrifuge for 10 min;

[0073] (4) Abandon the discard, add 10 μl of Buffer 1 resuscitation; add the following flow antibody, labeled mature B cells:

[0074] Anti-CD5 / Fitc (UCHT2, EBISCIENCE)

[0075] Anti-CD19 / PE (SJ25-C1, BD Pharmingen)

[0076] Anti-CD10 / APC (BC96, EBIoscience)

[0077] Anti-CD27 / PE-CY7 (O323, EBIoscience)

[0078] Anti-IgM / PE-CY5 (G20-127, BD Pharmingen)

[0079] (5) FACS SORTER sorted mature B cells (CD5-CD19 + CD10-CD27-IgM +), collected a single cell into a 96-well PCR plate, built-in 4 μl of cell lysa...

Embodiment 2

[0097] Example 2 Detection of antibody characteristics

[0098] 1, self-reactive detection

[0099] Antibody self-reactivity adopts clinical standard antinear antibody detection kit (Quanta Lite TM Anaelisa, Inova Diagnostics, Inc.) Perform a negative reaction sample, a low-positive reaction sample, a high-positive reaction sample in the detection kit. 4E10 is an HIV broad spectrum and antibody, also a self-reactive / multi-reactive antibody, and we have prepared 4E10 SCFV-Fc in an experiment for a positive control antibody. The test steps are as follows:

[0100] (1) Package: The ELISA board in the ANA kit has been carried out;

[0101] (2) Anti-anti - 50 μg / mL SCFV-Fc protein 50 μl, room temperature reaction for 2 h;

[0102] (3) Secondary antibody: Abandon the liquid, PBST was washed 3 times, add HRP-labeled sheep anti-human IgG-Fc antibody, and at room temperature 1 h;

[0103] (4) Selective: Abandon the liquid, PBST was washed 3 times, add TMB substrate solution, 100 μL per...

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PUM

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Abstract

The invention discloses an anti-HIV broad-spectrum neutralizing antibody, which comprises heavy chain variable regions CDR1, CDR2, and CDR3 with amino acid sequences shown in SEQ ID NO.1, 2, and 3 respectively; and SEQ ID NO. The light chain variable regions CDR1, CDR2, and CDR3 of the amino acid sequences shown in NO.5, 6, and 7. The invention also discloses nucleic acid encoding the antibody, a vector containing the nucleic acid, a host cell, an immunocoupling agent and a pharmaceutical preparation. The invention also discloses a method for producing HIV antibody and a screening method.

Description

Technical field [0001] The present invention belongs to the cellular immunology, the field of gene engineering, involves a broad spectrum and antibody of HIV. Background technique [0002] AIDS, Acquired Immunodeficiency Syndrome (AIDS) is immunodeflative disease caused by human immunodeficiency virus, HIV infection. The clinical manifestation of AIDS is infection, which causes the number of CD4 + T lymphocytes in the body to decrease, significantly affects the immune function of the body; in the late disease, CD8 + T lymphocytes will also decrease, and even finally destroyed, Loss resistance to disease capabilities, causing lesions of opportunistic infections, cancer, and central nervous systems, leading to AIDS patients to die for various composite infections. [0003] HIV broad spectrum neutralizing antibodies, because identifying HIV virus membrane proteins relatively conserved sites, which have little effect on HIV virus variation, which can resist most HIV virus strains, an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/10C12N15/13G01N33/569A61K39/42A61K45/06A61P31/18
CPCA61K39/42A61K45/06A61K2039/505A61P31/18C07K16/1045C07K2317/56C07K2317/565C07K2317/76G01N33/56988G01N2333/16G01N2469/10G01N2800/52A61K2300/00
Inventor 常宏刘松柏
Owner SUZHOU HEALTH COLLEGE
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