Solid-phase synthesis method of vMIP-II (viral macrophage inflammatory protein-II) protein

Abstract

The invention discloses a solid-phase synthesis method of vMIP-II protein. The steps are as follows: add dimethyldichlorosilane to the reaction bottle and ventilate overnight to silanize the reaction bottle, roughly wash with ethanol, place it at -80°C for cooling, and wash again Dry it, weigh the resin and place it in a reaction bottle, add DCM (dichloromethane) solution 3 times the volume of the resin, drain the liquid after swelling for 30 minutes, and then detect it on the polypeptide synthesizer according to the reaction of coupling amino acid and ninhydrin The cycle of deprotection with the α-amino group proceeds from the C-terminus to the N-terminus until all amino acid residues are coupled. After the coupling is completed, remove the reaction bottle and wash it with DCM, drain the resin and cut it with pre-cooled cutting solution, and purify the cut polypeptide by HPLC. The polypeptide was purified on a Waters600E high-pressure liquid chromatograph, and a Kromasil100-5C18 column was used for high-pressure reversed-phase chromatography, and the target peak polypeptide collected was the vMIP-II protein.

Description

technical field [0001] The invention relates to a chemical synthesis method of polypeptide, in particular to a solid-phase synthesis method of protein vMIP-II. Background technique [0002] Viral macrophage inflammatory protein-II (vMIP-II) is a viral gene product encoded by Kaposi's sarcoma virus (also known as human herpesvirus 8). The whole vMIP-II gene encodes 94 amino acids, and the mature protein is 74 amino acids, which has a homology with human MIP as high as 41%, of which 8 non-polar amino acids account for 40%, and have strong hydrophobicity. [0003] vMIP-II protein can not only play a role in the pathological response of immune inflammation by binding to chemokine receptors, but also can act on chemokine receptors on immune cells to regulate the function of immune cells by agonizing or antagonizing receptors , has more research and application value in HIV infection / AIDS and transplant rejection. [0004] The applicant has successfully used the E. coli prokaryo...

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Problems solved by technology

[0004] The applicant has successfully used the E. coli prokaryotic expression system to express vMIP-II, but due to the need to go through the process of vMIP-II inclusion body denaturation and renaturation, the purification cycle is long, the preparation cost is also high, and industrial production and product application are limited.

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