Xiaoyan Hu, Guangji Li, Ende Huang
School of Materials Science and Engineering, South China University of Technology, Guangzhou 510640, PR China.
Journal of chromatography. B, Analytical technologies in the biomedical and life sciences 2012 Jun 15L-histidine is a promising alternative to expensive protein ligands for the adsorption of IgG due to its high selectivity, no toxicity and low cost; while click chemistry can improve the reaction selectivity between the ligands and the support matrix under mild reaction conditions. Thus, using L-histidine as a ligand and original sepharose gel as a support, a novel immunoadsorbent possessing pseudo-biospecific affinity for IgG from human plasma, Sep-triazole-His, was designed and prepared according to the principle of Click-reaction between alkyne and azide functional groups; while both sepharose-based control samples Sep-His and Sep-PA were prepared by a conventional method using L-histidine and protein A as a ligand, respectively. The ligand density and IgG adsorption performance of Sep-triazole-His from human plasma were measured and evaluated. The influences of click chemistry on the preparation, structure and performance of sepharose-based immunoadsorbent were also investigated. The results indicate that the ligand density immobilized on Sep-triazole-His is 319.1 μmol/g sepharose gel, almost 4-fold as high as that on Sep-His; the IgG adsorption capacity of Sep-triazole-His from human plasma reaches 16.49 mg/g at pH 7.0, or increases 5.72-fold with respect to Sep-His, and does not decrease noticeably after being repeatedly used for 10 times; and Sep-triazole-His can exhibit high adsorption selectivity for IgG comparable to Sep-PA. The further studies prove that the 1,2,3-triazole ring in the spacer-arm of Sep-triazole-His, can facilitate the binding of IgG without non-specific adsorption. Copyright © 2012 Elsevier B.V. All rights reserved.
Xiaoyan Hu, Guangji Li, Ende Huang. Click chemistry: a route to designing and preparing pseudo-biospecific immunoadsorbent for IgG adsorption. Journal of chromatography. B, Analytical technologies in the biomedical and life sciences. 2012 Jun 15;899:96-102
PMID: 22664056
View Full Text