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Anti-human CD45RA rat immune globulin variable region gene and application

A technology of immunoglobulin and variable region, applied in anti-animal/human immunoglobulin, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, genetic engineering, etc.

Active Publication Date: 2012-05-09
杭州永申生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no systematic research on the antigen-targeted therapy at home and abroad.

Method used

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  • Anti-human CD45RA rat immune globulin variable region gene and application
  • Anti-human CD45RA rat immune globulin variable region gene and application
  • Anti-human CD45RA rat immune globulin variable region gene and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Nucleotide sequence and amino acid sequence of two genes of the present invention:

[0024] 1. Anti-human CD45RA mouse immunoglobulin heavy chain variable region gene (mAb ZCH-6-3A4VH 3A4 ) nucleotide sequence (SEQ ID NO 1) and amino acid sequence (SEQ ID NO 3).

[0025] 2. Anti-human CD45RA mouse immunoglobulin light chain variable region gene (mAb ZCH-6-3A4VL 3A4 ) nucleotide sequence (SEQ ID NO 2) and amino acid sequence (SEQ ID NO 4).

Embodiment 2

[0027] The ability of 3A4 to recognize antigen and its blocking effect on BD company's anti-CD45RA monoclonal antibody (clone name L48) were detected by flow cytometry inter-standard method ( figure 1 ), 1B in the figure is the flow cytometry diagram of the reaction between 3A4 and KG1a cell line, 1C is the flow diagram of the comparison with the anti-CD45RA monoclonal antibody of BD Company, and 1D is the flow diagram of the blocking experiment of 3A4 anti-CD45RA antibody. The results showed that the positive rate of 3A4 and KG1a cell reaction was 99.51%, and the MFI was 432.63, indicating that the activity of 3A4 antibody was good. figure 1 Peak 1 in E is the negative control, peak 2 is the image after blocking, and peak 3 is the image without blocking, indicating that 3A4 can completely block the reaction between BD’s anti-CD45RA antibody and KG1a, suggesting that the anti-CD45RA antibody and 3A4 recognize the same antigenic epitope.

[0028] Proceed as follows:

[0029] ...

Embodiment 3

[0036] Identification of 3A4 immunoglobulin subclasses: using flow cytometer inter-standard method, KG1a cell line as the reaction cell, 3A4 as the primary antibody, immunoglobulin subclass identification kit as the secondary antibody, and the detection of different immunoglobulins as the secondary antibody Anti-time positive rate and mean fluorescence intensity, see figure 2 , figure 2 In order to adopt the flow cytometry inter-standard method, KG1a cell line was used as the reaction cell, 3A4 was used as the primary antibody, and FITC-labeled goat anti-mouse immunoglobulin subclass antibody was used as the secondary antibody. The positive rate and average of different secondary antibodies were detected. Fluorescence intensity (MFI). The results showed that when IgG1 FITC was used as the secondary antibody, the positive rate was 98.48%, and the MFI was 332.85. When Kappa FITC was used as the secondary antibody, the positive rate was 99.02%, and the MFI was 252.20, indicati...

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Abstract

The invention provides an anti-human CD45RA rat immune globulin variable region gene, which comprises nucleotide sequences of SEQIDNO.1 and SEQIDNO.2 and encoded nucleotide sequences of SEQIDNO.3 and SEQIDNO.4. Research proves that anti-human CD45RA rat immune globulin variable region gene can effectively distinguish leukemia patients, leukemia cell strain CD34+CD38-CD123 stem cells and CD34+CD38+CD123 offspring leukemia cells, has the application prospects of leukemia targeted therapy, and can be used in preparing targeted therapy blood system malignant tumor medicines.

Description

technical field [0001] The invention belongs to biological technology, and mainly relates to anti-human CD45RA mouse immunoglobulin ZCH-6-3A4 (abbreviated as 3A4) (monoclonal antibody or monoclonal antibody) variable region gene and its use in the preparation of targeted drugs for treating hematological malignancies use. Background technique [0002] Hematological malignancy is a disease that seriously endangers human health. Before the 1970s, leukemia was considered an incurable disease, and its case fatality rate was almost 100%. With the in-depth research on the diagnosis and treatment of leukemia in various countries in the world, the continuous advent of new anti-tumor drugs and the implementation of hematopoietic stem cell transplantation, the prognosis of leukemia has been significantly improved. However, severe toxic and side effects are still the main obstacle to the successful clinical treatment of leukemia, and the lack of drug options for relapsed and drug-resis...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/13C07K16/28A61K39/395A61P35/02
Inventor 汤永民李思思
Owner 杭州永申生物科技有限公司
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