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Fully human anti-MAGEA1 all-molecule IgG antibody and application thereof

A fully human, DNA molecular technology, applied in the fields of applications, antibodies, antibody medical components, etc., to achieve good affinity and high specificity

Active Publication Date: 2016-11-09
秦皇岛未名健长幸医疗健康科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there is no report on the Chinese patent on the MAGE A1 antibody, and there are only related patents on MAGE A1 gene therapy

Method used

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  • Fully human anti-MAGEA1 all-molecule IgG antibody and application thereof
  • Fully human anti-MAGEA1 all-molecule IgG antibody and application thereof
  • Fully human anti-MAGEA1 all-molecule IgG antibody and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Preparation of fully human whole molecule IgG antibody against MAGEA1

[0030] 1) Using the MAGEA1 antigen in the human Fab phage library to undergo six rounds of "adsorption-elution-amplification" enrichment screening to obtain the anti-MAGEA1 Fab antibody, and then obtain the variable region sequence of the Fab antibody by PCR amplification and sequencing.

[0031] 2) Design primers according to the obtained antibody heavy and light chain variable region sequences.

[0032] 3) Amplify the heavy chain and light chain of the anti-MAGEA1 antibody.

[0033] Using the human Fab template prepared above, use the above-mentioned upstream and downstream primers for the heavy chain and light chain to amplify the heavy chain and light chain genes of the whole molecule human antibody, respectively.

[0034] (1)PCR

[0035] The reaction system is as follows:

[0036]

[0037] The reaction conditions are as follows:

[0038]

[0039] (2) 2% agarose gel electroph...

Embodiment 2

[0062] Example 2 Identification of Functional Activity of Anti-MAGEA1 Antibody

[0063] 1) ELISA

[0064] Coat ELISA 96-well plate with coating solution (0.1M carbonate buffer, pH9.6) MAGEA1 protein to 2 μg / mL, add 100 μL to each well, overnight at 4°C; PBST (PBS containing 0.5% Tween20) 5% skimmed milk -Wash buffer to block, incubate at 37°C for 2h; after washing with PBST 5 times, add 100μL PA21 antibody (2μg / mL initial concentration, 14 concentration gradient dilutions) to each well for 2h at 37°C; goat antibody diluted 1:4000 Add 100 μL / well of human secondary antibody to the well, incubate at 37°C for 1 hour; 100 μL / well of peroxidase substrate chromogenic solution, stop the reaction with 2M sulfuric acid after 10 minutes at room temperature, and use dual-wavelength 450nm / 690nm.

[0065] The result is as figure 2 shown by figure 2 It can be seen that the anti-MAGEA1 antibody can react with the MAGEA1 protein, and has a strong binding ability.

[0066] 2) Western b...

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Abstract

The invention discloses a fully human anti-MAGEA1 all-molecule IgG antibody. The fully human anti-MAGEA1 all-molecule IgG antibody comprises a light chain variable region and a heavy chain variable region, wherein the amino acid sequence of the light chain variable region is as shown in SEQ ID NO. 3, or the sequence is a conservative variant obtained by conservative mutation added, deleted, substituted and modified by one or more amino acids; and the amino acid sequence of the heavy chain variable region is as shown in SEQ ID NO. 4, or the sequence is a conservative variant obtained by conservative mutation added, deleted, substituted and modified by one or more amino acids. The invention further discloses DNA molecules, an expression vector, host cells and application of the all-molecule IgG antibody.

Description

technical field [0001] The present invention relates to the technical field of biological immunity, in particular to a fully human anti-MAGEA1 full-molecule IgG antibody, and also relates to the DNA molecule, expression vector, host cell and application of the full-molecule IgG antibody. Background technique [0002] Melanoma antigens (MAGE) are a group of tumor-associated antigens first discovered from melanoma. MAGE A1 (melanoma antigen A1) is actually a member of the MAGE gene family. Its encoded protein is found in a variety of tumor tissues and normal testicular tissues but not expressed in other normal tissues, so it is also called tumor-testis antigen. It is a tumor rejection antigen, which can be processed in cells to produce antigenic peptides, presented by HLA-IV molecules, recognized by autotoxic T lymphocytes, and undergoes antigen presentation and immune response to produce humoral immunity and cellular immunity. The immune response effect specifically kills th...

Claims

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Application Information

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IPC IPC(8): C07K16/18C12N15/13A61K39/395A61P35/00G01N33/574
CPCA61K39/00C07K16/18C07K2317/52C07K2317/56C07K2317/565C07K2317/73C07K2317/92
Inventor 朱进冯振卿许国贞刘振云唐奇熊四平
Owner 秦皇岛未名健长幸医疗健康科技有限公司
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