HSA (Human Serum Albumin) long-acting fusion protein and rapid assembly, separation and purification method thereof

Abstract

The invention relates to an HSA (Human Serum Albumin) long-acting fusion protein and a rapid assembly, separation and purification method thereof. The fusion protein comprises two polypeptide regions, wherein the first region is formed by a repetitive sequence of Exendin-4, the second region is formed by a sequence of the HSA, the C-end of the first region is connected with the N-end of the second region via glycine and serine, or the N-end of the first region is connected with the C-end of the second region via glycine and serine. The Exendin-4 is repeated for once to 4 times, and the repetitive sequence of the Exendin-4 is directly connected via glycine and serine. By adopting a Bglbrick method by the invention, an arbitrary number of Exendin-4 can be arbitrarily added on the N-end or C-end of the HSA, then high-purity fusion protein is acquired through expression and purification.

Description

technical field [0001] The present invention relates to a long-acting fusion protein of HSA (Human serum albumin, Human serum albumin) and its rapid assembly, separation and purification method, specifically the rapid assembly, separation and purification of HSA long-acting fusion protein by using the Bglbrick method, which belongs to the long-acting The technical field of fusion protein medicine. technical background [0002] The Biobrick / Bglbrick method is a different-scale DNA in vitro assembly method based on the idea of ​​ligation or aggregation. The birth of this method has accelerated the artificial construction of synthetic biological functional component libraries, biosynthetic pathways and even yeast chromosomes, and promoted metabolic engineering and genome editing. , Induced pluripotent hepatocytes and other fields of rapid development. [0003] The Biobrick / Bglbrick method is a method that relies on isotail enzymes. Isotail enzymes (such as BglII and BamHI, Xba...

AI Technical Summary

Problems solved by technology

The content of natural products is less, while the chemically synthesized Exendin-4 has disadvantages such as high cost, high price, many steps, low yield, and harmful chemicals involved in the synthesis process

The retention time of protein drugs in the body greatly affects the dosage and therapeutic effect of drugs. In recent years, scholars at home and abroad mainly extend the half-life of protein drugs from chemical modification and gene fusion. The most widely used chemical modification The modifier is PEG, which can prolong the half-life by several tens of times or even hundreds of times, but the immunogenicity of most proteins will be reduced, and the biological activity of proteins will also decrease to varying degrees

[0005] Human serum albumin (Human serum albumin, HSA) is the main protein component in plasma, and it is also the carrier of many endogenous factors and exogenous drugs, and plays a very important role in maintaining osmotic pressure and plasma volume in the body; under normal conditions, the kidney low clearance

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