Allogenic animal serum albumin as carrier protein for preparation of semiantigen-antibody

A technology of serum albumin and carrier protein, applied in the field of preparation of hapten antibodies, can solve the problems of influence, unfavorable screening of specific antibody titer monoclonal antibody, etc.

Active Publication Date: 2008-03-12
SHANGHAI KEXIN BIOTECH
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

This will have a very adverse effect on the titer of specific antibodies and the screening of monoclonal antibodies

Method used

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  • Allogenic animal serum albumin as carrier protein for preparation of semiantigen-antibody
  • Allogenic animal serum albumin as carrier protein for preparation of semiantigen-antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Mouse albumin as a hapten carrier for the preparation of FK506 monoclonal antibody

[0023] We used the fusion method of BALB / C mouse splenocytes and myeloma cells to prepare the monoclonal antibody of FK506 (trade name Prokof), FK506 is a macrolide antibiotic, composed of a semiketone group, Composed of a diketone group and 23 rings, 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 1. 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 used rivanol method to purify serum albumin of BALB / C homologous mice, obtained derivatives of FK506 by acid anhydride method, and then used EDC method to couple FK506 to homologous animal serum albumin.

[0024] 1. Purification of mouse albumin

[0025] 1) Collect a total of 5ml of mouse serum, centrifuge at 4000rpm / min for 15 minutes at 4°C, and ta...

Embodiment 2

[0063] Mouse albumin as hapten for preparation of BNP monoclonal antibody

[0064] 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.

[0065] The preparation method of serum albumin is the same as embodiment 1.

[0066] 1. The specific steps of coupling are as follows:

[0067] 1) Take 1mg of recombinant brain nasin (BNP), dissolve it in 1ml of double distilled water, and dissolve 10mg of self-made mouse serum albumin in 1ml of PBS (pH7.5) with a concentration of 0.1mol / L;

[0068] 2) shake and mix the hapten solution and the albumin solution;

[0069...

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Abstract

The present invention relates to a preparation method for a semi-antigen antibody. In details, the present invention discloses a preparation method for semi-antigen antibody by means of serum albumin of various animals as the carriers. According to the preparation method, first of all, semi-antigens of serum albumin of various animals are coupled into full antigens; and then the full antigens are used to generate antibodies. The serum albumin is protein which exists in the animal body since the embryo development. The immune cells already have immune no-response and the serum albumin is of no allotype or co-phenotype allele. The serum albumin does not vary from animal to animal and takes a large amount in the blood. Serum albumin can be easily obtained and the purifying process is simple. Therefore, the semi-antigen antibody prepared through the method is of higher value and better specificity.

Description

technical field [0001] The invention relates to a preparation method of hapten antibody, in particular to a method for preparing hapten antibody using the same animal serum albumin as a carrier. 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 presenting can not activate lymphocytes, but can be recognized by antibodies that have been produced, that is, immunoreactive but not immunogenic. To prepare small molecule hapten antibodies, the first step is to make small molecules with low molecular weight into larger molecules, and then prepare antibodies according to the method of c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18
Inventor 杨子义高景燕司徒维娜张晴妮李飞
Owner SHANGHAI KEXIN BIOTECH
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