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Anti-HIV broad-spectrum neutralizing antibody

An antibody and carrier technology, applied in the fields of cellular immunology and genetic engineering, can solve the problems of poor immunogenicity and virus mutation, and achieve the effect of good application prospects.

Active Publication Date: 2021-01-22
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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  • Anti-HIV broad-spectrum neutralizing antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] The preparation of embodiment 1 anti-HIV antibody

[0068] 1. Sorting of mature B cells

[0069] Buffer1: 500ml 1×PBS+2ml 0.5M EDTA+25ml 10%BSA (containing 2mM EDTA, 0.5%BSA)

[0070] (1) Collect 200 μl of peripheral blood from humanized BLT mice and add 200 μl of Buffer1;

[0071] (2) Add 10 times the volume of ACK lysing buffer (Fisher / BioWhittaker), incubate at room temperature for 10 minutes, and centrifuge at 1500 rpm for 10 minutes;

[0072] (3) Discard the supernatant, add 10ml Buffer 1 to wash the cells, and centrifuge at 1500rpm for 10min;

[0073] (4) Discard the supernatant, add 10 μl Buffer 1 to resuspend the cells; add the following flow cytometry antibodies to label mature B cells:

[0074] anti-CD5 / FITC (UCHT2, eBioscience)

[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) Sorting mature B cells (CD5-...

Embodiment 2

[0097] The detection of embodiment 2 antibody characteristics

[0098] 1. Self-reactivity detection

[0099] Antibody self-reactivity was detected using a clinical standard antinuclear antibody detection kit (QUANTA Lite TM ANAELISA, INOVA Diagnostics, Inc.) were tested, and negative reaction samples, low positive reaction samples, and high positive reaction samples were provided in the detection kit. 4E10 is an HIV broad-spectrum neutralizing antibody and also an autoreactive / polyreactive antibody. In the experiment, we prepared 4E10 scFv-Fc as a positive control antibody. The detection steps are as follows:

[0100] (1) Coating: the Elisa plate in the ANA kit has been coated with antigen;

[0101] (2) Primary antibody: Add 50 μl of 50 μg / ml scFv-Fc protein, react at room temperature for 2 hours;

[0102] (3) Secondary antibody: Discard the liquid in the well, wash 3 times with PBST, add HRP-labeled goat anti-human IgG-Fc antibody, and react at room temperature for 1 hour...

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PUM

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Abstract

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

Description

technical field [0001] The invention belongs to the fields of cellular immunology and genetic engineering, and relates to a broad-spectrum neutralizing antibody against HIV. Background technique [0002] AIDS, or acquired immunodeficiency syndrome (Acquired Immunodeficiency Syndrome, AIDS), is an immunodeficiency disease caused by human immunodeficiency virus (Human Immunodeficiency Virus, HIV) infection. The clinical manifestation of AIDS is that the number of CD4+ T lymphocytes in the immune system of the body is greatly reduced after infection, which significantly affects the immune function of the body; in the late stage of the disease, the CD8+ T lymphocytes will also decrease, and even be completely destroyed in the end. Loss of ability to resist diseases, causing opportunistic infections, cancer and central nervous system lesions, leading to death of AIDS patients due to various complex infections. [0003] HIV broad-spectrum neutralizing antibody, because it recogni...

Claims

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Application Information

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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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