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Preparation method of pepsinogen I paired monoclonal antibody

A monoclonal antibody, pepsinogen technology, applied in biochemical equipment and methods, anti-enzyme immunoglobulin, instruments, etc., can solve the problems of large workload and narrow screening range, and achieve good linearity and good application potential. Effect

Active Publication Date: 2018-08-14
BEIJING LEADMAN BIOCHEM
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has a large workload and relatively narrow screening range

Method used

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  • Preparation method of pepsinogen I paired monoclonal antibody
  • Preparation method of pepsinogen I paired monoclonal antibody
  • Preparation method of pepsinogen I paired monoclonal antibody

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Experimental program
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Embodiment 1P

[0037] The preparation method of embodiment 1PG I paired monoclonal antibody

[0038] 1. Experimental materials

[0039] SP 2 / 0 was purchased from the Cell Center of Shanghai Institute of Biochemistry and kept by the R&D Department of our company;

[0040] Balb / c mice were purchased from Shanghai Weitong Lihua Experimental Animal Center;

[0041] Fetal bovine serum: product of Hangzhou Sijiqing Company;

[0042] DMEM, HAT and HT: all are products of Invitrogen;

[0043] 50%PEG 1450 Solution: Sigma company product;

[0044] Other drugs are of analytical grade.

[0045] 2. Experimental method

[0046] 2.1 Immunization of Balb / c mice

[0047] Select 4-6 6-8 week old female Balb / c mice and immunize them with PG I antigen expressed by yeast.

[0048] The amino acid sequence of PG I (NP_001073276) is shown in SEQ ID No.1.

[0049] (1) The first fusion immunization of mice: BALB / c mice were immunized with yeast-expressed and purified PG I protein for a total of 3-5 times of ...

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Abstract

The invention discloses a preparation method of a pepsinogen I paired monoclonal antibody. The preparation method of the pepsinogen I paired monoclonal antibody comprises the following steps: (1) immunizing BALB / c mice with pepsinogen I protein as an antigen, and selecting immunized mice with high serum antibody titer for cell fusion, screening a high-titer positive cell line, preparing ascites, and purifying to obtain a monoclonal antibody; and (2) mixing the pepsinogen I protein with the monoclonal antibody obtained in the step (1) to prepare antigen-monoclonal antibody immune complex immunized BALB / c mice, selecting mice with high serum antibody titer for cell fusion, screening a positive cell line having no competitive inhibition effect with the monoclonal antibody of the step (1), andpurifying the secreted monoclonal antibody to obtain a monoclonal antibody paired with the monoclonal antibody of the step (1). The method effectively improves the probability of obtaining the pairedmonoclonal antibody of the pepsinogen I antigen.

Description

technical field [0001] The present invention relates to a preparation method of pepsinogen (PG) I paired monoclonal antibody, and the invention also relates to the paired monoclonal antibody of pepsinogen I prepared by the method and its hybridoma cell line, belonging to pepsinogen I Field of preparation of paired monoclonal antibodies. Background technique [0002] Pepsinogen (PG) is the precursor of pepsin. According to its biochemical properties and immunogenicity, it is divided into 2 subgroups. Components 1-5 have the same immunogenicity and are called pepsinogen I. Secreted by chief cells and mucous neck cells of fundic glands; components 6 and 7 are called pepsinogen II, in addition to being secreted by chief cells and mucous neck cells of fundic glands, mucous necks of cardiac glands and pyloric glands of gastric antrum Cells as well as the upper duodenum also produce pepsinogen II. [0003] Serum PG levels reflect the morphology and function of gastric mucosa in d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/40C12N5/20G01N33/573
CPCC07K16/40G01N33/573
Inventor 张晓杰熊宁
Owner BEIJING LEADMAN BIOCHEM
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