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cBIN1 antibody and application thereof

An antibody, cbin1 technology, applied in the field of immunity, can solve the problems of E-C decoupling cardiac function, decreased transfer ability, disturbance and other problems

Active Publication Date: 2021-05-14
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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  • cBIN1 antibody and application thereof
  • cBIN1 antibody and application thereof
  • cBIN1 antibody and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1: Synthesis of antigens for immunization

[0060] The antigen of the present invention is human Cardiac bridging integrator 1 exon 13 (cBIN1 exon13) corresponding protein sequence LRKGPPVPPPPKHTPSKEVKQEQILSLFEDTFVPEISVTTPSQ, the polypeptide used for immunization was synthesized by Jill Biochemical Co. Stearate (PEG4) conjugated keyhole limpet hemocyanin (KLH, keyhole limpet hemocyanin).

Embodiment 2

[0061] Example 2: Production of anti-human cBIN1 exon13 antibody

[0062] In order to obtain the murine anti-human cBIN1 exon13 antibody, the immunization strategies in Table 1.1 (Table 1) were used to immunize different strains of mice (Balb / c, Shanghai Lingchang Biotech; SJL, Beijing Weitong Lihua). The antigens used were as described in Example 1; the adjuvants include: complete Freund's adjuvant CFA (InvivoGen Company, catalog number vac-cfa-60), IFA (InvivoGen Company, catalog number vac-ifa-60). Three days after the final vaccination, the splenocytes of the immunized mice were fused with mouse myeloma cells SP2 / 0 using the polyethylene glycol method to obtain B cell fusions that could express antibodies and proliferate indefinitely in vitro, and were selected in HAT cultured in culture medium. The fused hybridoma cells were plated in a 96-well cell culture plate, and positive clones were selected through primary screening for two rounds of subcloning.

[0063] Table 1....

Embodiment 3

[0066] Example 3: Determination of the variable region sequence of the anti-human cBIN1 exon13 antibody

[0067] The hybridoma cells were collected by centrifugation, and each 5×10 6 ~10×10 6 Add 1ml TRIzol and 0.2ml chloroform to the cells, vibrate vigorously for 15 seconds, place at room temperature for 3 minutes, centrifuge to take the water phase, add 0.5ml isopropanol, leave at room temperature for 10 minutes, collect the precipitate, wash with ethanol and dry to obtain RNA. Add template RNA and primers into the ice-bath centrifuge tube, make the primers and templates paired correctly, then carry out the reverse transcription process, and then perform PCR amplification. Add 2.5 μl of dNTP / ddNTP mixture to each of the four microcentrifuge tubes, and incubate the mixture at 37°C for 5 minutes, and set aside. Add 1 pmol of PCR-amplified double-stranded DNA, 10 pmol of sequencing primers, 2 μl of 5× sequencing buffer to an empty microcentrifuge tube, add double-distilled wa...

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PUM

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Abstract

The invention relates to the technical field of immunity, in particular to a cBIN1 antibody and application thereof. Three CDR regions of a heavy chain of the cBIN1 antibody provided by the invention respectively have amino acid sequences as shown in SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3; and three CDR regions of a light chain respectively have amino acid sequences as shown in SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6. The antibody has good binding capacity with an antigen, and accurate detection of a target antigen can be realized by using the antibody to prepare a reagent.

Description

technical field [0001] The invention relates to the field of immune technology, in particular to cBIN1 antibody and its application. Background technique [0002] Heart failure is a group of complex clinical syndromes caused by abnormal changes in the structure and / or function of the heart caused by a variety of reasons, resulting in dysfunction of ventricular systolic and / or diastolic function. Heart failure is a severe manifestation or advanced stage of various heart diseases, with high mortality and rehospitalization rates. In recent years, data from various places have shown that the prevalence of heart failure in my country has increased significantly to 2% to 3%, and there are about 10 million current patients. [0003] The core pathological change of heart failure myocardium is the weakening of calcium ion transport ability, which limits the contractility of myocardium. The cause of the calcium ion transport disorder characteristic of heart failure is excitation-con...

Claims

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Application Information

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