A kind of biomimetic small peptide ligand-based affinity-enriched monolithic material and its preparation and application

Abstract

The invention belongs to the field of affinity enrichment integral materials and discloses an affinity enrichment integral material based on biomimetic small peptide ligands, preparation and an application. A substrate integral material is prepared from monomer GMA, a crosslinking agent, a pore-foaming agent and an initiator through a mixed reaction; then a metal chelator solution is poured into the substrate material, a heating reaction is performed, then a metal inorganic salt solution is poured for a reaction, so that metal ions are chelated and immobilized to the substrate material, then ahistidine-tagged small peptide ligand solution is poured for a reaction, and the affinity enrichment integral material based on the biomimetic small peptide ligands is obtained. Small peptides are taken as affinity ligands of the integral material, compared with the commonly used biomacromolecular ligands such as protein A / G, antigens or target proteins and the like, the small peptides have the advantages of being low in cost, stable in chemical property, free of immunogenicity and long in service life and containing no biological impurities, and conformation of an antibody protein cannot bedamaged due to quite mild elution conditions.

Description

technical field [0001] The invention belongs to the field of affinity-enrichment monolithic materials, in particular to an affinity-enrichment monolithic material based on bionic small peptide ligands, as well as its preparation and application. Background technique [0002] Monoclonal antibody drugs have become a hot field in the pharmaceutical industry due to their many advantages such as strong targeting, high specificity, and low toxic and side effects. Antibody drugs accounted for seven of the top ten drugs sold globally in 2014, and took the top three [Nature Reviews Drug Discovery, 2015, 14(2): 83-83; Nature Reviews Drug discovery, 2014, 13(8): 577 -587.]. According to the latest forecast of PMR, the world's top market research company, the global antibody drug market will exceed US$140 billion by 2020. It is worth noting that in the production process of monoclonal antibody drugs, the cost of downstream monoclonal antibody purification accounts for about 50-80% of ...

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Problems solved by technology

Synthetic biomimetic ligands are usually designed and synthesized using protein A / L / G as a template, with good stability, selectivity and affinity, and are suitable for the enrichment and purification of mAbs in cell culture supernatants, but for those containing a large amount of highly homologous IgG, etc. In vivo samples of heteroproteins, its specific selectivity is still insufficient [Bioanalysis, 2016, 8 (8): 847-856.], and the ligands such as dyes have the disadvantages of toxicity and poor biocompatibility [Methods, 2012, 56 (2 ):116-129.]

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