The prognosis prediction model can be used for purifying hydrophobic cyclic peptide ligands of human immunoglobulin G

Abstract

The invention provides a hydrophobic cyclic peptide ligand capable of being used for separating and purifying human immunoglobulin G. The hydrophobic cyclic peptide ligand is obtained through a computer simulation bionic design method, and the bionic design method takes an amino acid sequence Ser383-Asn389 on a CH3 binding region on a SpA and hIgG-Fc fragment as a simulation system. Hydrophobic cyclic peptide ligands which can be specifically combined with hIgG are determined through LeDock molecular docking, FlexX molecular docking and Amber molecular dynamics simulation screening, and the cyclic peptide ligand chromatographic medium is adopted to perform purification research on hIgG from a human serum sample, so that purity as high as 96% and recovery rate as high as 92% can be obtained; the hydrophobic cyclic peptide ligand sequence is -[HWG-C-AKTE]-, -[YF-C-WRHE]-, [-WV-C-LHHYF]-, -[PYF-C-TIE]- and -[FY-C-DEHL]-, wherein the peptide ligand with the best effect is -[HWG-C-AKTE]-.

Description

technical field [0001] The present invention relates to the technical field of purification of human immunoglobulin G, which adopts computer simulation technology to bionically design an affinity peptide ligand library of human IgG, and then uses affinity chromatography test to verify the purification effect of peptide molecules on human IgG, which belongs to the field of bionic design Computer simulation and downstream protein separation and purification technology field. Background technique [0002] At present, antibodies are widely used in medical diagnosis and disease treatment. In 2006, the global sales of more than 20 kinds of antibody drugs approved by FDA exceeded 17 billion US dollars. Among them, IgG is the main antibody component in serum, accounting for about 75% of serum Ig. , is also the class of antibodies with the greatest demand. The preparation of antibody drugs is usually completed by precipitation, ion exchange chromatography, hydrophobic interaction ch...

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Problems solved by technology

[0004] The research group of Professor Sun Yan from Tianjin University used 6 key residues of SpA: F132, Y133, H137, E143, R146 and K154 to construct a simulation system, and used molecular docking and molecular dynamics simulation to screen the peptide library. The purity of human IgG can be obtained at The linear octapeptide molecules FYWHCLDE and FYTHCAKE with a recovery rate of more than 88% and a recovery rate of more than 65% have good purification effects, but there is still a long way to go before commercial application, which is based on the relationship between protein A and IgG. Hydrophobic interaction model is used to simulate, but the obtained polypeptide sequence is based on electrostatic interaction and hIgG binding. It is well known in the art that electrostatic interaction is not specific adsorption, and it is easy to adsorb other biomass molecules. The purity of the obtained protein is not high, and it is difficult to commercialize use

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