Antibody cBIN1 and application thereof

An antibody, cbin1 technology, applied in the field of immunity, can solve problems such as decline, E-C decoupling of cardiac function, and decreased metastatic ability

Active Publication Date: 2021-04-02
THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In heart failure, in addition to a series of remodeling changes in the morphology of the transverse canal, the damaged LTCC migrates to the transverse canal, and the process of forming a binary complex with RyRs is also disturbed, Ga 2+ Decreased metastasizing capacity, leading to E-C uncoupling and decreased cardiac function

Method used

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  • Antibody cBIN1 and application thereof
  • Antibody cBIN1 and application thereof
  • Antibody cBIN1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059]Example 1: Synthesis of antigen for immunization

[0060]The antigen of the present invention is human Cardiac Bridging Integrator 1 Excmine 13 (Cbin1 EXON13) corresponding protein sequence LRKGPPPPPPPPPPPPSkevkqeqilslfedTFVPEISVTTPSQ, and the polypeptide-proof biochemical company used for immunization is synthesized by Guo Hui, in order to improve antigen-immunogenicity, the polypeptide C-terminal passed two Stearate (PEG4) coupling keylet blood blue protein (KLH, Keyhole Limpet Hemocyanin).

Embodiment 2

[0061]Example 2: Production of anti-human source Cbin1 EXON13 antibody

[0062]In order to obtain a mouse anti-human Cbin1 EXON13 antibody, immunization strategy (Table 1) in Table 1.1 was used to immunize different lines of mice (Balb / C, Shanghai Ling Chang; SJL, Beijing Virong Lihua). The antigen used is as described in Example 1; adjuvant includes: complete Frego adjuvant CFA (INVOGEN, INVOGEN VAC-CFA-60), IFA (INVOGEN, IQ VAC-IFA-60). After three days of enduring, the spleen cells of immunized mice were fused using polyethylene glycol cells using polyethylene glycol cells, which can express the antibody and in vitro, which can be blended in vitro, and in HAT selection Culture in the medium. The fused hybridoma cells were placed in a 96-well cell culture plate and 2 rhythmium subclon was selected by primary screening.

[0063]Table 1. Immune strategy

[0064]

[0065]

Embodiment 3

[0066]Example 3: Determination of variable region of anti-human source Cbin1 EXON13 antibody

[0067]Centrifugation collection hybridoma cells, every 5 × 106~ 10 × 106The cells were added to 1 ml of Trizol and 0.2 ml of chloroform, strenuous oscillated for 15 seconds, and 0.5 ml of isopropanol was added at room temperature, and 0.5 ml of isopropyl alcohol was added. After 10 minutes at room temperature, the ethanol was mixed and dried to give RNA. Templates RNA and primers are added to the ice bath, which makes the primer and template correctly paired the reverse transcription process, and PCR amplification. 2.5 μL of DNTP / DDNTP mixture was added to 4 microcencular tubes, and the mixture was given 37 ° C for 5 min, spare. Add 1 pmol of PCR amplified double-stranded DNA, 10Pmol sequencing bracket, 2 μl 5 × sequencing buffer, add double steamer to total volume 10 μL, 1min, 1 min, 4 ° C 10000 g from centrifugation 10S. 2 μl of pre-cooling label mixture (DCTP, DGTP, DTTP 0.75 μmol / L), ...

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Abstract

The invention relates to the technical field of immunization and particularly relates to an antibody cBIN1 and application thereof. Three CDR regions of a heavy chain of the antibody cBIN1 provided bythe invention separately have amino acid sequences represented by SEQ ID NO: 1, 2 and 3; and three CDR regions of a light chain of the antibody cBIN1 separately have amino acid sequences representedby SEQ ID NO: 4, 5 and 6. The antibody has good binding ability with an antigen, and reagents prepared from the antibody can achieve accurate detection on target antigens.

Description

Technical field[0001]The present invention relates to the field of immunization, and more particularly to CBIN1 antibodies and their applications.Background technique[0002]Heart failure is a variety of abnormal changes caused by a variety of reasons, so that the ventricular shrinkage and / or diastolic function occurs, thereby causing a set of complex clinical syndrome. Heart failure is a serious manifestation or late stage of various heart diseases, and mortality and re-hospitalization rates are high. According to data from all over the past few years, the prevalence of my heart is significantly increased to 2% to 3%, and the patient is about 10 million.[0003]The core pathological changes of cardiac failure is weakened, so that the contraction of myocardium is limited. The cause of calcium ion transmission disorder caused by heart decline is excitation-contraction (E-C) disabled and non-synchronized calcium release. Under physiological conditions, the membrane potential is polarize...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18C12N15/13G01N33/68
CPCC07K16/18G01N33/6893C07K2317/565C07K2317/56C07K2317/567C07K2317/92C07K2317/24G01N2800/325G01N2333/47
Inventor 赵元周康
Owner THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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