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Anti-her2 affibody, and switchable chimeric antigen receptor using same as switch molecule

A technology of chimeric antigen receptors and affimers, which is applied in the direction of antibody medical components, anti-receptors/cell surface antigens/cell surface determinant immunoglobulins, antibodies, etc., which can solve the problem of uncontrollable chimeric antigen receptor T cells Activation and expansion, application restrictions and other issues

Pending Publication Date: 2021-12-21
ABCLON
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Despite impressive success in early-stage clinical trials, the inability to control the activation and expansion of existing chimeric antigen receptor T cells (CAR-T cells) in vivo remains limited

Method used

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  • Anti-her2 affibody, and switchable chimeric antigen receptor using same as switch molecule
  • Anti-her2 affibody, and switchable chimeric antigen receptor using same as switch molecule
  • Anti-her2 affibody, and switchable chimeric antigen receptor using same as switch molecule

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0150] Example 1. Development of affiBody in HER2

Embodiment 1-1

[0151] Example 1-1. Screening by Panning (PANNING)

[0152] A cloning body with HER2 specific binding was screened from the affinity library (Library) using HER2-ECD-FC proteins. Further, five kinds of complexes were screened by confirming the binding clones binding to the cells expressing HER2.

[0153] The affinity library is used in phage morphological rescue by using VSCM13 auxiliary phage (Helper Phage). First, the number of library phages used to combine the antigen is 10 13More than the 4 rounds of Panning Rounds. The method used is as follows: The strategy of selectively screening the high phage of affinity, with the gradual increase in the number of times, gradually reduces the amount of antigen (10 μg, 5 μg, 2 μg, 1 μg), gradually increase the cleaning (WASH) ) Times (3 times, 5 times, 7 times, 10 times).

[0154] The binder pHAGE obtained in each wheel obtained by the infection (INFECTION) obtained by an enzyme-linked immunosorbent assay (ELISA) is used to confirm whet...

Embodiment 1-2

[0158] Example 1-2. Confirmation of the combination of parentures

[0159] The five kinds of affinity clones (Cloning) were screened into a Fc binding form (ZB-Fc) to confirm the degree of binding to HER2 protein and the degree of binding to HER2 expression cells. The enzyme-linked immunosorbent assay was carried out by the following method: 5 ZB-Fc purified from 60 nm was diluted to 1 / 5, and the above-mentioned ZB-Fc was coated with HHER2-ECD-His. 7 points were treated on the culture plate of the protein, and the second antibody (Anti-Higg-Fc-HRP (INVITROGEN company, H10007)) was treated, 3, 3 ', 5,5'-Tetracycline is used. After the acid-linked immunosorbent assay reader is used to determine OD using an enzyme-linked immunosorbent assay reader. 450 Value, seeking EC via Graph Prism 50 value( image 3 ,Table 2).

[0160] Table 2

[0161] distinguish ZQAA1 ZQAA7 ZQAA8 ZQAA11 ZQAA22 EC 50 (nm)

3.6 68.5 1.8 6.9 N / D

[0162] In order to confirm the degree...

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PUM

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Abstract

The present invention relates to an anti-HER2 affibody, and a switch molecule comprising a cotinine-coupled anti-HER2 affibody. The anti-HER2 affibody of the present invention, when coupled with cotinine, reacts to positive cell lines in which HER2 is expressed if treated together with Cot-sCART, so as to induce the activity of immunocytes, and thus can be effectively used as a switch molecule for an sCART therapeutic agent.

Description

Technical field [0001] Under the support of the Ministry of Korea Industrial University, the present invention has been achieved through the topic number 20002893. The research management specialized agencies of the above topics are the Korea Industrial Technology Evaluation Administration, the research institution name is "Biological Industry Core Technology Development," research topics For the development of HER2 targeted ovarian cancer therapeutic agents using innovative CART technology, the competent authority is ABCLON (strain) company, and the study period is from October 01, 2018 June 30, 2021. [0002] The present application claims priority to Korean Patent Application No. 10-2020-0047025, Ji 17, 2020, which is incorporated herein by reference. [0003] The present invention relates to an "anti-HER2 pin and a switchable chimeric antigen receptor that uses it as a switching molecule." Background technique [0004] Although impressive success is obtained in the clinical t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/32C07K16/28C07K19/00A61K39/00A61K31/4439A61K47/68A61P35/00
CPCC07K16/32C07K16/2863C07K14/7051C07K16/44A61K39/001119A61K31/4439A61K47/6849A61K47/6851A61P35/00C07K2317/92C07K2319/03C07K2319/33C07K2317/565C07K2317/622C07K2317/73C07K19/00A61P37/02A61K47/6855A61K47/6803C07K2318/20A61K39/001106A61K2039/505A61K2039/5156A61K2039/572C12N5/0636C12N2510/00C07K2318/00A61K47/545C07K2319/02
Inventor 金起贤李炫宗李种瑞
Owner ABCLON
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