A Novel Humanized Fab Phage Display Vector

Abstract

The invention discloses a new type of human Fab phage display carrier, which is connected with a human light chain conserved region sequence: CL in front of the XbaI restriction site of the vector, and a human heavy weight is connected in front of the vector SpeI restriction site. Chain conserved region sequence: CH1, a 1000bp insert is connected after the SacI restriction site of the vector: VL-insert, and a 914bp insert is connected before the vector XhoI restriction site: VH-insert. The present invention relates to the technical field of phage display. The novel humanized Fab phage display vector directly connects the human heavy chain CH1 region and the light chain CL region into the vector, and adds enzyme cleavage sites at both ends, thereby reducing the number of The workload improves the quality and diversity of the library construction. A large fragment of about 1Kb is introduced into the variable region of the heavy chain and the variable region of the light chain, thereby reducing the uncut plasmid when cutting the gel to recover the linear plasmid fragment. Contamination is simple and effective, and the correct rate of constructing vectors for human Fab phage library is improved.

Description

Technical field [0001] The present invention involves the field of phage display technology, which is a new type of human FAB phage display carrier. Background technique [0002] The phage display technology is to use the phage carrier or phage carrier to clone the polypeptide or protein encoding gene fragment into the appropriate position of the pharmaceutical case protein structure gene, so that the exogenous polypeptide or protein and the specific phage clothing protein are integrated, displayed in A technology on the surface of the phage, the polypeptide or protein displayed through this technology can maintain a relatively independent spatial structure and biological activity to a certain extent, which is conducive to the specific identification and combination of target molecules, and ensures that the molecule genotypes and Expressive unity. [0003] In recent years, phage technology has become a binding site for detecting protein space, exploring the interaction between re...

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Problems solved by technology

[0007] Aiming at the deficiencies of the prior art, the present invention provides a new type of human Fab phage display vector, which solves the problem of large workload when connecting variable region fragments, and the fact that overlap PCR easily leads to sequence errors, and at the same time over-amplifies VH. The increase may cause the problem of the decline of gene diversity in the variable region

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