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Anti-human MSLN (mesothelin) monoclonal antibody

A monoclonal antibody, antibody technology, applied in the direction of antibodies, anti-animal/human immunoglobulin, anti-tumor drugs, etc., can solve the problem of no public report on antibody screening scheme, no nano antibody, etc., to shorten the screening process , Low immunogenicity, good effect of antibody heterogeneity

Active Publication Date: 2020-01-17
ZHEJIANG BLUE SHIELD PHARM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As far as antibodies targeting MSLN are concerned, there are no public reports on the development of nanobodies, and there are no public reports on the screening of antibodies targeting MSLN based on phage display technology.

Method used

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  • Anti-human MSLN (mesothelin) monoclonal antibody
  • Anti-human MSLN (mesothelin) monoclonal antibody
  • Anti-human MSLN (mesothelin) monoclonal antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0032] (1) Phage display antibody library panning

[0033] 1) Host TG1 activation: Prepare mini arar medium plate [1×M9 salt, 2% glucose, 2mM MgSO 4 , 0.1 mM CaCl 2, 1mM vitamin B1], streak overnight culture TG1 in a 37 ℃ incubator.

[0034] 2) Coating: Dilute human MSLN protein (purchased from Acro Biosystem) in 0.1M NaHCO at a concentration of 50ug / ml 3 (pH8.6), 1.5ml was added to the immunotube, and coated overnight at 4°C. After removing the coating solution, incubate with blocking solution [0.01M PBS (pH7.4), 2% skimmed milk powder] for 1 hour, and wash 4 times with 0.01M PBS (pH7.4) after the end.

[0035] 3) Combination: take 10 11 Add pfu phage virus particles to 1.5ml MPBS [0.01M PBS (pH7.4), 2% skimmed milk powder] in the immunotube, and incubate at room temperature for 1 hour. After removing the liquid, wash 8 times with PBST [0.01M PBS (pH7.4), 0.1% Tween-20], and then wash 8 times with 0.01M PBS (pH7.4).

[0036] 4) Elution: Rinse the immunotube repeatedly w...

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Abstract

The invention discloses an anti-human MSLN (mesothelin) monoclonal antibody, which is a specific high-affinity nano antibody developed by taking human MSLN as a target and based on a phage display technology. Compared with the prior art, the invention allows the anti-human MSLN monoclonal antibody to be successfully prepared, which is good in specificity, high in affinity and capable of binding tohuman MSLN expressed on the cell surface, and the anti-human MSLN monoclonal antibody is a potential drug for tumor immunotherapy.

Description

technical field [0001] The invention relates to an anti-human MSLN monoclonal antibody, belonging to the technical field of monoclonal antibodies. Background technique [0002] Mesothelin (MSLN) is a 40KD glycoprotein located on the cell surface, anchored to the cell membrane by glycosylphosphatidylinositol. The mesothelin gene encodes a 69kDa precursor protein, which is hydrolyzed into two chains by furin (paired basic amino acid protease, furin)-like invertase, and the membrane-bound protein of about 40KD at the C-terminal is the mature mesothelial protein The N-terminal 30KD fragment called megakaryocyte-promoting factor (MPF) falls off and is released outside the cell. Both MPF and membrane anchor MSLN are N-glycosylated, MPF can promote the formation of megakaryocyte clones in vitro, membrane anchor MSLN can interact with MUC16, and plays an important role in the process of cell adhesion, so currently targeted therapy is Membrane-anchored MSLN is selected as the targe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/30C12N15/13G01N33/577G01N33/574A61K39/395A61P35/00
CPCC07K16/30C07K16/005G01N33/577G01N33/57484A61P35/00C07K2317/622C07K2317/569C07K2317/92A61K2039/505
Inventor 张军锋郭志刚
Owner ZHEJIANG BLUE SHIELD PHARM CO LTD
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