Click chemistry: a route to designing and preparing pseudo-biospecific immunoadsorbent for IgG adsorption.
L-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.
School of Materials Science and Engineering, South China University of Technology, Guangzhou 510640, PR China.
Xiaoyan
X
Hu
Guangji
G
Li
Ende
E
Huang
D000327:Adsorption; D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D057930:Click Chemistry / Q000295:instrumentation / Q000379:methods*; D006639:Histidine / Q000737:chemistry; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry* / Q000302:isolation & purification; D007164:Immunosorbents / Q000737:chemistry*; D007700:Kinetics; D008024:Ligands; D013045:Species Specificity
D007074:Immunoglobulin G; D007164:Immunosorbents; D008024:Ligands; D006639:Histidine