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Anti-human Legumain protein monoclonal antibody, hybridoma cell strain LGMN-2 and applications thereof

一种杂交瘤细胞株、LGMN-2的技术，应用在抗酶免疫球蛋白、生物化学设备和方法、仪器等方向，能够解决应用还属空白、没有检测方法等问题，达到好特异性的效果

Inactive Publication Date: 2019-04-30
李伟 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] As a new biological indicator, Legumain has no clinical detection method for it at present, and its application in monitoring the occurrence and development of tumors is still blank

Method used

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  • Anti-human Legumain protein monoclonal antibody, hybridoma cell strain LGMN-2 and applications thereof
  • Anti-human Legumain protein monoclonal antibody, hybridoma cell strain LGMN-2 and applications thereof
  • Anti-human Legumain protein monoclonal antibody, hybridoma cell strain LGMN-2 and applications thereof

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Embodiment 1

[0031] Embodiment 1, prepare the monoclonal antibody of anti-human Legumain protein

[0032] 1. Preparation of Legumain antigen

[0033] 1) Obtaining the sequence of the Legumain gene

[0034] The gene encoding Legumain is located on human chromosome 14 (14q32.12), and its sequence number in the NCBI database is CCDS: CCDS9904.1. The plasmid pLEXSY-sat2-hLGMN carrying the human Legumain gene was donated by Dr. Elfriede Dall from the University of Salzburg, Austria. The non-patent literature that has recorded the eukaryotic expression plasmid pLEXSY-sat2-hLGMN is E. Dall and H. Brandstetter, Activation of legumain involves proteolytic and conformational events, resulting in a context-and substrate-dependent activity profile., Acta Crystallogr Sect F Struct Biol Crystal Commun. 2012 Jan 1;68(Pt 1):24–31.

[0035] 2) Construction and identification of recombinant plasmids

[0036] In order to make human Legumain secreted outside the cell after expression, a tissue-type plasmin...

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Abstract

The invention discloses an anti-human Legumain protein monoclonal antibody, a hybridoma cell strain LGMN-2 and applications thereof. The name of provided hybridoma cell strain is LGMN-2, and the preservation number is CGMCC No.12995. The provided anti-human Legumain protein monoclonal antibody is secreted by the hybridoma cell strain LGMN-2 with a preservation number of CGMCC No.12995. The provided monoclonal antibody has a high titer, and the maximal indirect method titer can reach 1:2.56*10<8>. There is no cross reaction between the monoclonal antibody and Legumain proteins: Cathepsin B, Cathepsin H, and Cathepsin L; and the specificity is good.

Description

technical field [0001] The invention relates to the technical field of biomedical engineering, in particular to a monoclonal antibody against human Legumain protein, a hybridoma cell line LGMN-2 and uses thereof. Background technique [0002] Legumain (C13) (EC 3.4.22.34), also known as asparaginyl endopeptidase (AEP), is a lysozyme cysteine ​​protease and a new member of the cysteine ​​protease C13 family. The human Legumain gene is located on chromosome 14, encoding a polypeptide chain composed of 433 amino acid residues, which becomes a biologically active mature enzyme after deglycosylation, and has 83% homology with mice. Human legumain protein has 3 forms, namely 56kD zymogen precursor, 46kD and 36kD mature enzyme. The active catalytic center of Legumain is a catalytic duplex composed of histidine (His) and cysteine ​​(Cys), which is necessary for Legumain to exert its catalytic activity and is also a necessary structure for every C13 family member. [0003] Compared...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/40G01N33/577G01N33/573
CPCC07K16/40G01N33/573G01N33/577G01N2333/96466
Inventor 李伟
Owner 李伟
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