Novel human-derived Fab phage display vector

Abstract

The invention discloses a novel human-derived Fab phage display vector, and relates to the technical field of phage display. According to the novel human-derived Fab phage display vector, a human-derived light chain conserved region sequence CL is linked before a vector XbaI restriction enzyme cutting site, a human-derived heavy chain conserved region sequence CH1 is linked before a vector SpeI restriction enzyme cutting site, a 1000bp insertion fragment VL-insert is linked behind a vector SacI restriction enzyme cutting site, and a 914bp insertion fragment VH-insert is linked before a vectorXhoI restriction enzyme cutting site. According to the novel human-derived Fab phage display vector, a human-derived heavy chain CH1 region and light chain CL region are directly linked into the vectors, restriction enzyme cutting sites are added at the two ends of the vectors, so that the workload is reduced, and the quality and diversity of library construction are improved; and large fragmentsof about 1 Kb are each introduced into a heavy chain variable region and a light chain variable region, so that the contamination of uncut plasmids when linear plasmid fragments are recycled by cutting bales is reduced, simpleness and efficiency are achieved, and the accuracy of constructing vectors for human-derived Fab phage libraries is improved.

Description

technical field [0001] The invention relates to the technical field of phage display, in particular to a novel human Fab phage display carrier. Background technique [0002] Phage display technology uses phagemid vectors or phage vectors to clone polypeptide or protein coding gene fragments into appropriate positions of phage coat protein structural genes, so that foreign polypeptides or proteins can be fused with specific phage coat proteins and displayed on A technology on the surface of phage, through which the polypeptide or protein displayed can maintain a relatively independent spatial structure and biological activity to a certain extent, which is conducive to the specific recognition and binding of target molecules, and ensures the molecular genotype and Expressive unity. [0003] In recent years, phage technology has become an effective tool for probing the spatial structure of proteins, exploring the binding sites of interactions between receptors and ligands, and...

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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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