Anti-human MSLN monoclonal antibody and application thereof

A monoclonal antibody and antigen technology, applied in applications, antibodies, anti-animal/human immunoglobulins, etc., can solve the problem that the antibody screening scheme has not been publicly reported, save development time and cost, shorten the screening process, High affinity effect

Active Publication Date: 2020-06-30
NANJING BLUE SHIELD BIOTECH 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 fully human ScFv antibodies, 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 monoclonal antibody and application thereof
  • Anti-human MSLN monoclonal antibody and application thereof
  • Anti-human MSLN monoclonal antibody and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0030] Below in conjunction with specific example, further illustrate the present invention.

[0031] (1) Phage display antibody library panning

[0032] 1) Host TG1 activation: Prepare mini agar medium plate [1×M9 salt, 2% glucose, 2mM MgSO 4 , 0.1mM CaCl 2 , 1mM vitamin B1], streak overnight culture TG1 in a 37 ℃ incubator.

[0033]2) Coating: Dilute human MSLN protein (purchased from Acro Biosystem) in 0.1M NaHCO at a concentration of 50ug / ml 3 (pH8.6), take 1.5ml into the immunotube, and coat overnight at 4°C. After removing the coating solution, incubate with blocking solution [0.01M PBS buffer (pH7.4), 2% skimmed milk powder] for 1 hour, and wash 4 times with 0.01M PBS buffer (pH7.4) after the end.

[0034] 3) Combine: take 10 11 Add pfu phage virus particles to 1.5ml MPBS solution [0.01MPBS buffer (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 solution [0.01M PBS bu...

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Abstract

The invention discloses an anti-human MSLN monoclonal antibody and an application thereof, and is a specific and high-affinity single-chain antibody developed by taking human MSLN as a target based ona phage display technology. Compared with the prior art, the anti-human MSLN monoclonal antibody is successfully prepared, the antibody is good in specificity and high in affinity and can be combinedwith human MSLN expressed on the surface of a cell, and the anti-human MSLN monoclonal antibody is a potential drug for tumor treatment.

Description

technical field [0001] The invention belongs to the technical field of monoclonal antibodies, in particular to an anti-human MSLN monoclonal antibody and its application. Background technique [0002] Mesothelin (MSLN) is a 40KD glycoprotein located on the cell surface, which is 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-ancho...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/30C12N15/13G01N33/574A61K39/395A61P35/00
CPCA61P35/00C07K16/005C07K16/30C07K2317/24C07K2317/92G01N33/57484
Inventor 张军锋郭志刚
Owner NANJING BLUE SHIELD BIOTECH CO LTD
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