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human cd70 antigen binding polypeptide

A technology for binding peptides and antigens, applied in the field of human CD70 antigen-binding polypeptides, can solve the problems of high engineering risk, expensive, time-consuming, etc., and achieve the effects of easy passage through blood vessels, high affinity and good specificity

Active Publication Date: 2020-06-16
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 the current level of technological development is concerned, antibody humanization is still a challenging task, and the technical obstacles are not small. The project is risky, time-consuming, labor-intensive, and expensive

Method used

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  • human cd70 antigen binding polypeptide
  • human cd70 antigen binding polypeptide
  • human cd70 antigen binding polypeptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0031] (1) Phage display peptide library panning

[0032] 1) Activation of host strain ER2738 (purchased from NEB): Prepare a sterile LB plate containing 20ug / ml tetracycline (1g / 100ml peptone, 0.5g / 100ml yeast powder, 0.5g / 100ml NaCl), and culture ER2738 overnight by streaking in a 37°C incubator.

[0033] 2) Coating: Dilute human CD70 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, use blocking solution [0.1M NaHCO 3 (pH8.6), 5mg / ml BSA] for 1 hour, and then washed 4 times with TBST [50mM Tris-Cl (pH7.5), 150mM NaCl, 0.1% Tween-20].

[0034] 3) Combination: take 10 11 Pfu phage virus particles (purchased from NEB) were added to the immune tube containing 1.5ml TBST and incubated for 1 hour. After removing the coating solution, wash 4 times with TBST.

[0035]4) Elution: wash the immunotube repeatedly with 1ml eluent [0.2M g...

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Abstract

The invention discloses a human CD70 antigen binding polypeptide. The human CD70 antigen binding polypeptide with higher specificity and higher affinity is developed with human CD70 as a target. Compared with the prior art, the human CD70 antigen binding polypeptide (fusion protein) is prepared successfully, the polypeptide has good specificity and higher affinity and can bind human CD70 expressedon cell surfaces, and the anti-human CD70 monoclonal antigen binding polypeptide is a potential drug for tumor immunotherapy.

Description

technical field [0001] The invention relates to a human CD70 antigen-binding polypeptide and belongs to the technical field of antigen-binding polypeptides. Background technique [0002] CD70 molecule, Tumor Necrosis Factor Receptor Superfamily 7 Factor (TNFSF7). The length of the polypeptide chain is 193 a.a., and the molecular weight is 21.1KD. It is a type II transmembrane glycoprotein (the soluble form has not yet been seen), and the 155 amino acids at the C-terminal are located outside the membrane. Disulfide bonds are formed between the first, third, second, and fourth cysteines outside the cell to maintain the spatial structure of the extracellular part. Usually it is present on the membrane as a homotrimer. [0003] In normal tissues, the cellular expression profile of CD70 is narrow, mainly expressed in B cells in germinal centers (to induce antibody secretion) and T cells in local lymphoid tissues, and expressed at a low level in thymic medullary epithelial cells...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/85C12N5/10G01N33/68G01N33/574A61K39/00A61P35/00
CPCA61K2039/505A61P35/00C07K14/70575C07K2319/02C07K2319/21C07K2319/30C12N15/85G01N33/574G01N33/6854G01N2333/70575
Inventor 张军锋郭志刚
Owner ZHEJIANG BLUE SHIELD PHARM CO LTD
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