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A kind of limulus peptide-antibody fusion protein and its preparation method

A technology of fusion protein and limulus peptide, applied in the direction of anti-animal/human immunoglobulin, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, chemical instruments and methods, etc., can solve the problem of antibacterial activity and other issues to achieve the effect of low toxic and side effects

Active Publication Date: 2018-04-24
TIANJIN LANRY BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

2. In terms of the utilization of limulus peptide, the above patents only stay in the application of crops and livestock and poultry breeding industry
3. The knowledge of the above-mentioned patents on the activity of limulus peptide still remains on its antibacterial activity

Method used

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  • A kind of limulus peptide-antibody fusion protein and its preparation method
  • A kind of limulus peptide-antibody fusion protein and its preparation method
  • A kind of limulus peptide-antibody fusion protein and its preparation method

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preparation example Construction

[0133] A method for preparing a limulus peptide-antibody fusion protein, using genetic engineering methods to modify the limulus peptide gene at the genetic level, and linking the limulus peptide gene T to the coding antibody unit directly or through the linking peptide gene LU encoding the linker unit The 3' end or the 5' end of the heavy or light chain of the gene M, forming (T-LU) N –M method, construct recombinant expression plasmid, express its fusion protein through host cell,

[0134] The preparation method comprises steps as follows:

[0135] (1) Using genetic engineering methods to connect the limulus peptide gene through the linker gene or directly to the antibody gene, and connect it to the expression vector to obtain a recombinant expression plasmid, the specific steps are as follows:

[0136] ①Using overlap extension PCR technology, using the anti-CD19Fab L chain with the sequence SQ20 as the template, through two PCRs, the G 4 The S gene, the T1 gene and the st...

Embodiment 1

[0163] 1. Obtain the light chain variable region gene (VL) and heavy chain variable region (VH) gene of anti-CD19 antibody:

[0164] The sequences of the light chain variable region gene (VL) and the heavy chain variable region (VH) gene of the anti-CD19 antibody are from the patent: Anti-CD19 engineering antibody for targeting lymphocytic leukemia cells and its use (publication number CN 1775808A).

[0165] The light chain variable region (VL) gene of the anti-CD19 antibody, that is, the nucleic acid sequence of anti-CD19VL is SQ24.

[0166] The heavy chain variable region (VH) gene of the anti-CD19 antibody, that is, the nucleic acid sequence of anti-CD19VH is SQ25.

[0167] The above two gene sequences were obtained by synthetic methods. At the same time, in order to facilitate the construction of the expression vector and the connection of the variable region and the constant region, a restriction site Mlu I and a protective base were introduced at the 5' end of the VL ge...

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Abstract

Provided are a tachyplesin-antibody fusion protein and preparation method thereof. The fusion protein comprises at least one tachyplesin unit, and the gene of a coded tachyplesin unit is directly connected to the 3' end or 5' end of a gene of a light chain or a coded antibody heavy chain, or is connected thereto via a gene of a coded linker peptide.

Description

technical field [0001] The present invention relates to the fields of oncology, pathogenic microbiology and biopharmaceuticals. The present invention relates to a limulus peptide-antibody fusion protein and its preparation method. The fusion protein containing limulus peptide and the preparation method thereof provide an excellent candidate drug for the targeted therapy of tumor and pathogenic microorganism infection. Background technique [0002] Limulus peptide is derived from Limulus blood cell extract and is a polypeptide containing 17-18 amino acid residues. Its amino and carboxyl terminals are basic amino acids, and the carboxyl terminal also has a unique arginine-amide structure. Four Cys form 2 disulfide bonds to maintain the limulusin peptide to fold into a hairpin structure. At present, 5 kinds of limulus peptides have been found, tachyplesin I and tachyplesin II are found in blood cells of Limulus orientalis, tachyplesin III is found in blood cells of Limulus pol...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/70
CPCC07K7/08C07K16/28C07K19/00
Inventor 何永胜苑庆华韩钊辛鹤林臧丹戎李彦彩李萌任卫新
Owner TIANJIN LANRY BIO TECH
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