A kind of ii type autoimmune liver lkm-1, lc-1 composite quality control product and its preparation method and application
A LKM-1, LC-1 technology, applied in the directions of botanical equipment and methods, biochemical equipment and methods, applications, etc., can solve the problems of batch difference, high cost, low yield, etc., and achieve cost reduction and matrix effect. Small, waste-reducing effect
Active Publication Date: 2022-04-08
四川携光生物技术有限公司
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[0011] The purpose of the present invention is to provide a type II autoimmune liver LKM-1, LC-1 composite quality control product and its preparation method, to solve the problem of high cost, low yield, and batch quality of existing LKM-1 and LC-1 antibody quality control products. big difference problem
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Embodiment 1
[0044] A kind of preparation method of type II autoimmune liver LKM-1, LC-1 composite quality control product, comprising the following steps:
[0045] S1. Obtain the variable region gene sequence of the mouse LKM-1 antibody and the variable region gene sequence of the mouse LC-1 antibody respectively, as shown in SEQ ID No.1 and SEQ ID No.2 respectively;
[0046] The process of obtaining the variable region gene sequence is as follows:
[0047] Recombinant antigen LKM-1 and recombinant antigen LC-1 obtained splenocytes expressing mouse LKM-1 antibody and mouse LC-1 antibody respectively by immunizing mice several times; splenocytes were extracted and fused with Sp2 / 0 cells, screened, It was verified that cell lines that can express LKM-1 and LC-1 monoclonal antibodies at high levels were obtained respectively; the cell lines were further injected intraperitoneally into mice, and the ascites was extracted and purified to obtain the corresponding monoclonal antibodies, and the ...
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The invention discloses a type II autoimmune liver LKM-1, LC-1 composite quality control product and its preparation method and application. The preparation method includes the following steps: S1, respectively obtaining the variable region gene of the mouse LKM-1 antibody Sequence and the variable region gene sequence of the mouse LC-1 antibody are shown in SEQ ID No.1 and SEQ ID No.2 respectively; S2, insert a rigid Linker between SEQ ID No.1 and SEQ ID No.2 Connect, and insert the EcoR I restriction site at one end, and insert the other end sequence into the Nco I restriction site to synthesize the complete gene sequence, as shown in SEQ ID No.3; S3, introduce the complete gene sequence obtained in step S2 into pFUSE‑ In the hIgG1-Fc2 vector, prepare a recombinant plasmid containing both the mouse LKM-1 antibody and the mouse LC-1 antibody variable region; S4, transfect the recombinant plasmid obtained in step S3 into mammalian cells, and obtain a recombinant plasmid containing both the mouse LKM-1 antibody and the variable region of the mouse LC-1 antibody; 1 antibody and a recombinant antibody to the variable region of the mouse LC‑1 antibody. The invention solves the problems of high cost, low output and large batch-to-batch difference of existing LKM‑1 and LC‑1 antibody quality control products.
Description
technical field [0001] The invention relates to the technical field of in vitro diagnosis, in particular to a type II autoimmune liver LKM-1, LC-1 composite quality control product and its preparation method and application. Background technique [0002] Autoimmune liver disease (AIH) is a multi-etiological disease. Viruses, bacteria, and chemical substances induce genetically sensitive individuals to participate in the inflammatory disease process, but the defect of immune regulation function is often the basic cause of AIH. [0003] According to different immune serological markers, AIH is currently divided into 4 types. Type I AIH, formerly known as classic type or lupus type, is the most common type of the disease, accounting for about 60% to 80% of all AIH. It is characterized by: ANA and (or) SMA positive, hyperglobulinemia, more common in women (accounting for 71%), effective hormone therapy, easy to be complicated by extrahepatic autoimmune diseases such as thyroidit...
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IPC IPC(8): C07K16/18C12N15/13C12N15/85G01N33/96G01N33/68G01N33/543
CPCC07K16/18C12N15/85G01N33/96G01N33/6854G01N33/6893G01N33/54326C07K2317/56C12N2800/22G01N2800/085
Inventor 刘欣潘敬梅秦枫黄敬双张小飞颜松周玥张伟万文琴鲜静
Owner 四川携光生物技术有限公司