Human apolipoprotein AV monoclonal antibody and application thereof

A monoclonal antibody and apolipoprotein technology, applied in the direction of anti-animal/human immunoglobulin, biochemical equipment and methods, instruments, etc., can solve the problems such as no reports of ApoAV monoclonal antibodies, and achieve a linear detection range Large and stable effect

Inactive Publication Date: 2009-11-11
湖南远泰生物技术有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, there is no repor

Method used

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  • Human apolipoprotein AV monoclonal antibody and application thereof
  • Human apolipoprotein AV monoclonal antibody and application thereof
  • Human apolipoprotein AV monoclonal antibody and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 , Antigen preparation

[0030] Using EX-W0804-B01 (purchased from GeneCopoeia) as a template, using F.primer and R.primer as primers, perform PCR amplification, and then connect the obtained PCR product to the PQE30 vector to obtain recombinant strains, and ferment them And purify to obtain the desired antigen, the specific process is as follows:

[0031] 1. Primer design

[0032] According to the sequence of the target gene Genbank Acc#: NM_052968.3, design primers F.primer and R.primer, and their sequences are as follows:

[0033] F. primer: CGC GGATCC GCACGGAAAGGCTTCTGGGA;

[0034] R. primer: CCC AAGCTT TCAGGGGTCCCCCAGATGGCT.

[0035] Among them, the underlined marks in the primers F.primer and R.primer are their BamH I and Hind III restriction sites, respectively.

[0036] 2. PCR amplification

[0037] Using EX-W0804-B01 (purchased from GeneCopoeia) as a template and using F.primer and R.primer as primers, PCR amplification was carried out. The ...

Embodiment 2

[0047] Example 2 , Preparation of monoclonal antibodies

[0048] Using human serum lipoprotein AV as an antigen, Balb / c mice were used as immunization objects to prepare monoclonal antibody cells, and the obtained stable positive monoclonal antibody cells were cultured and purified to obtain human serum lipid Monoclonal antibody to protein AV, the specific process is as follows:

[0049] 2.1. Animal immunity

[0050] With Balb / c mice (purchased from Shanghai Experimental Animal Center, Chinese Academy of Sciences) as the immunization object, the human serum lipoprotein AV prepared in Example 1 was used as the antigen, and the animal was immunized according to the immunization scheme in Table 1, wherein, the first -4 times of immunization by subcutaneous immunization, and the fifth time by tail vein injection for booster immunization, and 4 days after immunization, the immunized Balb / c mice were sacrificed, and the splenocytes were harvested for use in cells fusion.

[005...

Embodiment 3

[0059] Example 3 , human serum lipoprotein detection

[0060] Serum samples were collected from 505 cases of outpatient physical examination in the Second Xiangya Hospital of Central South University from June to July 2006, including 259 males and 246 females, aged 20-76 (average 44±12) years old; diabetes, hyperlipidemia, hyperlipidemia were excluded Blood pressure, kidney disease, tumor and other immune system diseases, and TG≤2.4mmol / L, TCHO≤5.6mmol / L in blood lipid test results. In the morning, 2ml of blood was drawn on an empty stomach and placed in a quick-clotting tube, placed at room temperature for 2 hours, then centrifuged at 3000r / min, and the serum was taken for later use.

[0061] With the monoclonal antibody of the human serum lipoprotein AV prepared in embodiment 2, carry out westernblot or ELISA detection according to the following scheme:

[0062] 3.1, western blot detection

[0063] The obtained monoclonal antibody was used as the primary antibody, and HR...

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Abstract

The invention provides a human apolipoprotein AV monoclonal antibody and an application thereof. The human apolipoprotein AV monoclonal antibody is obtained by syncretizing a splenic cell and a myeloma cell of a human apolipoprotein AV immunized mouse and then cultivating the syncretized cell. The monoclonal antibody can be used for detecting human apolipoprotein AV in human serum. The Apo AV monoclonal antibody can be used for detecting Apo AV in human serum. When the Apo AV monoclonal antibody is used for detecting the Apo AV, the linear detection scope is wide, ApoAI, ApoB and Lp(a) in the serum do not influence a detection result, and the detection result has good stability, therefore, the Apo AV monoclonal antibody can be used for preparing corresponding detection products so as to detect the Apo AV concentration in the human serum, determine the high risk group of coronary heart disease as early as possible and prevent coronary heart disease in time and cure the coronary heart disease in the early period.

Description

technical field [0001] The present invention belongs to the field of biotechnology, specifically relates to a monoclonal antibody, more specifically relates to a human apolipoprotein AV monoclonal antibody and its application. Background technique [0002] Apolipoprotein AV is a new apolipoprotein discovered by Pennacchio et al. in 2001. It is located at the -27kb position downstream of ApoAIV. It is highly homologous to the base sequence of mouse ApoAV and human ApoAIV. The expressed protein is adjacent to The gene product ApoAIV also has a high degree of homology, so it is named ApoAV. [0003] According to studies, ApoAV is related to blood lipid levels, especially closely related to triglyceride levels, and triglyceride is an independent risk factor for coronary heart disease. lipoprotein. Therefore, the preparation of ApoAV monoclonal antibodies for the determination of ApoAV concentration in human serum is of great significance for early screening of high-risk groups...

Claims

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Application Information

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IPC IPC(8): C07K16/18C12N5/18C12P21/08G01N33/577
Inventor 张涛吴力军刘光耀田方艳赵水平胡敏彭道泉
Owner 湖南远泰生物技术有限公司
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