Preparation for self-protein used as carrier protein for hapten-antibody

A carrier protein and hapten technology, applied in the direction of anti-animal/human immunoglobulin, etc., can solve the problems of unfavorable antibody titer and specificity, influence, etc.

Inactive Publication Date: 2008-04-23
SHANGHAI KEXIN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This cross-linking method is relatively simple, but the disadvantage is that new protein structures are introduced, unwanted epitopes appear, and a

Method used

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  • Preparation for self-protein used as carrier protein for hapten-antibody
  • Preparation for self-protein used as carrier protein for hapten-antibody

Examples

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Effect test

Embodiment 1

[0024] Example 1 Preparation of FK506 polyclonal antibody

[0025] FK506 is a macrolide antibiotic, consisting of a semiketone group, α, β diketone group and 23 rings, and its molecular formula is C 44 h 69 NO 12 ·H 2 O, the molecular weight is 822. Its structural formula is shown in Figure 2. It alone cannot be used as an antigen to immunize animals to obtain specific antibodies, and needs to be coupled with a carrier protein to obtain immunogenicity. We obtained derivatives of FK506 by anhydride method, and then coupled with autologous serum albumin by EBC method. Specific steps are as follows:

[0026] 1) Dissolve 20 mg of FK506 in 5 ml of pyridine, add 50 mg of glutaric anhydride, and react at 80°C for 4 hours at reflux, and use silica gel thin-layer chromatography to detect the progress of the reaction, and stop the reflux reaction when the reaction is nearly complete;

[0027] 2) solvent pyridine is removed by distillation under reduced pressure to obtain FK506 gl...

Embodiment 2

[0047]Example 2 BNP is coupled to serum albumin using a bifunctional cross-linking agent

[0048] The amino acid sequence of BNP (B-type natriuretic peptide, brain natriuretic peptide) is SSKMYQGSGCPGFKMDRLSSSSGLGCKVLRRR from the amino terminal to the carboxyl group. It is a peptide containing 32 amino acid residues. The relative molecular mass of BNP is 3.46×10 3 , it is not sufficient as a complete antigen to immunize animals, and needs to be conjugated to a carrier protein to obtain immunogenicity. Conjugate with serum albumin using a bifunctional cross-linker method. Specific steps are as follows:

[0049] 1) Dissolve 1mg of brain nasin (BNP) in 1ml of double distilled water; 10mg of self-made rabbit serum albumin, dissolve in 1ml of PBS (pH 7.5) with a concentration of 1mol / L;

[0050] 2) shake and mix the hapten solution and the rabbit serum albumin solution;

[0051] 3) Add 1 ml of 0.25% (volume ratio) glutaraldehyde dropwise while stirring. Stir for 5-10 minutes af...

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Abstract

The present invention relates to hapten antibody preparing process, and is especially the process of first preparing complete antigen through coupling animal's autologous protein to hapten and subsequent preparing antibody through immunizing animal with the complete antigen. The antibody preparing process has no introduced foreign protein, body produced antibody targeting to the small molecular weight compound and small peptide hapten, and no antibody to autologous protein. Therefore, the prepared hapten antibody prepared with autologous protein as carrier protein has high valence and high specificity.

Description

technical field [0001] The invention relates to a preparation method of a hapten antibody, in particular to a method for preparing an animal autologous protein as a carrier protein for a hapten antibody. Background technique [0002] Haptens are molecules that have the ability to bind antibodies but are unable to stimulate the body to produce antibodies. Antigens are composed of three necessary elements of macromolecules with heterogeneity, structural complexity and a certain molecular weight. Although haptens are heterogeneous and structurally complex, their molecular weight is not large enough to be recognized by immune cells alone in the body And presented so can not activate lymphocytes, produce antibodies, but can be recognized by antibodies that have been generated, that is, immunoreactive but not immunogenic. To prepare small-molecule hapten antibodies, the first step is to make the low-molecular-weight small-molecule haptens larger molecules, and then prepare antibo...

Claims

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Application Information

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IPC IPC(8): C07K16/18
Inventor 杨子义司徒维娜高景燕徐骥贾世香
Owner SHANGHAI KEXIN BIOTECH
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