Induced expression and purification method of human-derived insulin-like growth factor binding protein 1 in pichia pastoris, and preparation and application of related antibodies

Abstract

The invention relates to an induced expression and purification method of a human-derived insulin-like growth factor binding protein 1 in pichia pastoris. A nucleotide sequence optimized by codons and used for coding the human-derived insulin-like growth factor binding protein 1 is connected with an HIS tag label, then is constructed into a pichia pastoris induced expression vector, next is transferred into pichia pastoris, and is subjected to induced expression; the expression product is subjected to affinity chromatography to obtain a purified human-derived insulin-like growth factor binding protein 1. The invention also relates to related antibodies and an application thereof in preparation of diagnostic kits for premature rupture of fetal membranes. The induced expression and purification method of the human-derived insulin-like growth factor binding protein 1 in pichia pastoris is skillful in design, and thus the human-derived insulin-like growth factor binding protein 1 can be efficiently expressed; and with the human-derived insulin-like growth factor binding protein 1 as an antigen, a high-specific polyclonal antibody and a high-specific monoclonal antibody for recognition of different IGFBP-1 epitopes are obtained, thereby laying the foundation for application of multiple downstream detection kits, and being suitable for large-scale popularization and application.

Description

technical field [0001] The present invention relates to the technical field of protein expression, purification and antibody preparation, in particular to the technical field of induced expression, purification and antibody preparation of protein in Pichia pastoris, specifically refers to the production of a human insulin-like growth factor binding protein 1 in Pichia pastoris Induced expression and purification methods, related antibody preparation and application. Background technique [0002] Human insulin-like growth factor-binding protein 1 (IGFBP1) is one of the important endocrine hormones discovered in the human body in the 1980s. It is composed of a group of soluble proteins and is divided into six members including IGFBP1-6. It binds to insulin-like growth factor (IGF) with high affinity, but different IGFBPs have different biological effects after binding to IGF. The full-length cDNA of IGFBP1 was reported for the first time in 1988. The main biological significa...

AI Technical Summary

Problems solved by technology

Human insulin-like growth factor-binding protein 1 has complex glycosyl groups and a specific spatial conformation, and the bacterial expression system cannot glycosylate IGFBP-1 protein due to the limitations of prokaryotes, which may also affect the normal folding of its spatial conformation

On this basis, the detection antibody is single, and there is still much room for improvement in detection sensitivity

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