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Covalent immobilization of antibodies for the preparation of immunoaffinity chromatographic supports.

SEP SCI TECHNOL. 2016-04; 
LA Mejíamanzano, J Gonzálezvaldez, K Mayolodeloisa, EJ Escalantevázquez, M Ritopalomares.
Products/Services Used Details Operation
... GenScript. (2015) THE PEG Antibody (Mouse). //www.genscript.com/peg_antibody_mab. html. View all references). ... No. A01795) was purchased from Gen Script (NJ, USA) and anti CD-133 antibody was kindly donated by Dr. Richard C. Willson of the University of Houston. ...

摘要

Immunosorbents in immunoaffinity chromatography (IAC) are prepared by immobilizing expensive antibodies without guidelines for ensuring the best coupling efficiencies, and avoiding low binding capacities. Covalent immobilization of antibodies on N-hydroxysuccinimide (NHS)-activated Sepharose 4 Fast Flow resin was optimized using human IgG via full factorial design with incubation times (4, 9, 14, 19 and 24 h), temperatures (4°C and 20°C) and coupling reaction buffers (sodium bicarbonate and triethanolamine). The best coupling efficiency (CE) (83.4 ± 8.7%) was reached with triethanolamine buffer, 14 h and 4°C. Comparison of antibody isotypes (IgG or IgM) by a nested factorial analysis suggested that antibodi... More

关键词

Coupling efficiency (CE), IgG, IgM, immunoaffinity chromatography (IAC), optimization