Anti-alanyl-trna synthetase single chain antibodies and methods of making the same

The preparation of anti-alanyl-tRNA synthetase single-chain antibodies using phage display technology solves the antibody acquisition problem, enables the preparation of high-titer antibodies, and is suitable for in vitro diagnostic kits, showing broad application potential.

CN118184791BActive Publication Date: 2025-11-21SHARETRY BIOTECH CO LTD
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Patent Information

Application Number
CN202410434828.1
Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Filing Date
2024-04-11
Publication Date
2025-11-21
Estimated Expiration
2044-04-11

AI Technical Summary

Technical Problem

There are currently no reports on single-chain antibodies against alanyl-tRNA synthetase, making it difficult to obtain high-titer antibodies for the diagnosis and treatment of antisyntheticase syndrome.

Method used

Single-chain antibodies against alanyl-tRNA synthetase were prepared using phage display technology. The DNA sequences, including the variable regions of the heavy and light chains, were used to amplify ScFv using RT-PCR, PCR, and overlap extension PCR. Combined with phage display screening, a high-titer antibody library was obtained.

Benefits of technology

This technology enables the production of antibodies with large antibody library capacity, simple operation, low cost, and high titer, laying the foundation for in vitro diagnostic kits and showing broad application prospects.

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Abstract

The application provides an anti-alanyl-tRNA synthetase single-chain antibody and a preparation method thereof, the anti-alanyl-tRNA synthetase single-chain antibody comprises a heavy chain variable region and a light chain variable region, the DNA sequence of the heavy chain variable region is shown as SEQ ID NO:1, and the DNA sequence of the light chain variable region is shown as SEQ ID NO:2. After healthy mice are immunized by using the antigen prepared in the laboratory, the antibody titer is qualified, the spleen is taken to extract RNA, and the ScFv is obtained through the modes of RT-PCR, PCR and overlap extension PCR; after being connected with a pCANTAB5e vector, phage antibody library is obtained through electric transformation; through multiple rounds of screening, the antibody sequence with high titer is obtained; the application has the advantages of large antibody library capacity, relatively simple operation, low cost and the like, lays a foundation for further application in an in-vitro diagnostic kit, and has a wide application prospect.
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