Pepsinogen II monoclonal antibody and application thereof

A monoclonal antibody, antigen-antibody technology, applied in immunoglobulins, anti-enzyme immunoglobulins, instruments, etc., can solve the problems of low antibody activity, poor affinity, and susceptibility to lipoprotein interference, avoiding interference and low requirements. Effect

Active Publication Date: 2021-06-29
重庆艾生斯生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide anti-PGII monoclonal antibody, to solve the problems of low activity, poor affinity and easy to be interfered by lipoproteins of existing antibodies, and to realize high-precision and high-sensitivity PGII detection

Method used

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  • Pepsinogen II monoclonal antibody and application thereof
  • Pepsinogen II monoclonal antibody and application thereof
  • Pepsinogen II monoclonal antibody and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1: Expression and purification of recombinant PGII

[0043] The human PGII gene sequence was cloned into a prokaryotic expression vector to construct a prokaryotic expression plasmid, and the expression plasmid was transformed into Escherichia coli BL21 and cultured on an LB plate containing antibiotics. Single clones were picked and inoculated into LB medium. After culturing until the OD value of the bacterial solution reached 0.6, IPTG was added to induce protein expression. After 16 hours of induction, the bacterial liquid was collected by centrifugation. The supernatant was collected by centrifugation after the cells were crushed, and purified by ion exchange and molecular sieve to obtain the antigenic protein.

Embodiment 2

[0044] Example 2: Mouse immunization and antibody detection

[0045] Five female BALB / c mice of SPF grade aged 6-8 weeks were selected, and Freund's complete adjuvant and PGII protein at a concentration of 2 mg / ml were mixed and emulsified in equal volumes. The emulsified antigen was immunized with 6-8 week-old SPF grade female BALB / c mice, and each mouse was injected with 40 μg of antigen protein by sole injection or back subcutaneous injection. Two weeks after the completion of the primary immunization, the antigenic protein was mixed with Freund's

[0046] The incomplete adjuvant was mixed and emulsified, and each mouse was injected with 40 μg of antigenic protein by plantar injection or back subcutaneous injection. Two weeks later, blood was collected through the tail vein, the supernatant was collected by centrifugation and the serum titer was detected by ELISA. Immunize once every two weeks and test the serum titer. After the second immunization, the serum titer after...

Embodiment 3

[0047] Example 3: Production and purification of monoclonal antibodies

[0048] Two groups of 6-8 week BALB / c mice were selected, and 500 μL paraffin oil was injected intraperitoneally to suppress the immune response of the mice. One week after the injection, 0.5 ml of PGII hybridoma cells were intraperitoneally injected into a group of mice, and the number of cells was about 1×10 6 quantity. Ascites collection began two weeks later. The collected ascites was subjected to ammonium sulfate precipitation and protein A affinity purification to obtain the target antibody.

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Abstract

The invention relates to the technical field of biology, in particular relates to a pepsinogen II monoclonal antibody and application thereof, and discloses a group of pepsinogen II monoclonal antibodies which can be used in cooperation, which can be applied to different methodologies, accurate measurement of PGII is achieved, interference of other impurities can be avoided, and high-specificity binding is achieved. The monoclonal antibody provided by the invention can assist in research and development of medical diagnostic reagents, and has great significance.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to pepsinogen II monoclonal antibody and application thereof. Background technique [0002] Pepsinogen (Pepsinogen or PG) is the precursor of pepsin, which is divided into pepsinogen I (PGI) and pepsinogen II (PGII) according to the difference in physical and chemical properties and immunogenicity, and is mainly secreted by the gastric mucosa. Pepsinogen has no digestive enzyme activity, and after being secreted into the gastric cavity, it is activated into pepsin with digestive activity under the action of gastric acid. In addition to its main function in the digestive tract, a small amount of pepsinogen will pass through the gastric mucosa and enter the blood circulation. The content of pepsinogen in serum can reflect the level of gastric juice secretion and the pathological changes of gastric mucosa. The content of PGII in normal human serum is 11.9±4.2ng / ml. Studies have pointed o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/40G01N33/577G01N33/573G01N33/574
CPCC07K16/40C07K2317/565C07K2317/92G01N33/573G01N33/57446G01N33/57488G01N33/577G01N2333/96477G01N2800/062G01N2800/065
Inventor 易维京蒋会会蔡方琴赵忠颢汤定斌温路新
Owner 重庆艾生斯生物工程有限公司
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