Anti-human GPC3 monoclonal antibody

A monoclonal antibody, antibody technology, applied in the direction of antibodies, anti-animal/human immunoglobulins, anti-tumor drugs, etc., can solve the problems of no nano antibody research and development, no public reports, etc. The combined effect of high and low immunogenicity

Active Publication Date: 2019-12-31
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 GPC3 are concerned, there are no public reports on the development of nanobodies, and there are no public reports on the screening of antibodies targeting GPC3 based on phage display technology.

Method used

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  • Anti-human GPC3 monoclonal antibody
  • Anti-human GPC3 monoclonal antibody
  • Anti-human GPC3 monoclonal antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0033] (1) Phage display antibody library panning

[0034] 1) Host TG1 activation: Prepare mini argar medium plate [1×M9 salt, 2% glucose, 2mM MgSO4, 0.1mM CaCl 2 , 1mM vitamin B1], streak overnight culture TG1 in a 37 ℃ incubator.

[0035] 2) Coating: Dilute human GPC3 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.

[0036] 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).

[0037] 4) Elution: Rinse the immunotube repeatedly wit...

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Abstract

The invention discloses an anti-human GPC3 monoclonal antibody which is a specific high-affinity nanometer antibody developed by using human GPC3 as a target on the basis of a phage display technology. Compared with the prior art, the anti-human GPC3 monoclonal antibody is successfully prepared; the antibody has high specificity and high affinity, and can be combined with the human GPC3 expressedon the cell surfaces; and the anti-human GPC3 monoclonal antibody belongs to potential medicine for tumor immunotherapy.

Description

technical field [0001] The invention relates to an anti-human GPC3 monoclonal antibody, belonging to the technical field of monoclonal antibodies. Background technique [0002] Glypican 3 (GPC3) is a heparan sulfate proteoglycan on the surface of the cell membrane, composed of a core protein (GPC), heparan sulfate (HS) and glycosylated phosphatidylinositol (GPI) A carbohydrate-lipid-protein complex formed by covalently linking three parts. HS is connected to the serine residue of GPC through the oligosaccharide chain, and the GPC protein is anchored on the surface of the cell membrane through the GPI through the Asp554 site. The HS connection site is the two serine residues at the C-terminus of GPC, so that the HS chain is close to the cell membrane. [0003] The core protein is composed of 580 a.a., the molecular weight is about 70KD, and the furin protease cleavage site is in the middle, which can be cut by furin from between Arg358 and Cys359 to form a 40KD N-terminal su...

Claims

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Application Information

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IPC IPC(8): C07K16/30C12N15/13C12N15/85C12N5/10G01N33/574A61K39/395A61P35/00
CPCA61K2039/505A61P35/00C07K16/005C07K16/303C07K2317/622G01N33/574G01N33/577
Inventor 张军锋郭志刚
Owner NANJING BLUE SHIELD BIOTECH CO LTD
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