[Production and characterization of the single chain antibodies against human alpha2b-interferon]

Abstract

A combinatorial library of single-chain antibodies (ScFv) from mice immunized with human alpha2b interferon (hIFN-alpha2b) was constructed. For obtaining of phage display antibodies the DNA fragments of ScFv were cloned into phagemid vector pCANTAB-5E and rescued from Escherichia coli cells by infection with bacteriophage M13. Bacterial clones synthesizing specific ScFv against hIFN-alpha2b were isolated by several rounds of affinity selection of phage library. After isolation and affinity purification of ScFv-IFN from bacteria cells a high ability to binding of the hIFN-alpha2b has been shown. The sequencing of isolated ScFv DNA and analysis of the data obtained have been carried out. The data of expression stability of obtained E. coli producers such as some features of ScFv expression are also discussed.