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Comparison of antibody functionality using different immobilization methods
Author(s) -
Danczyk R.,
Krieder B.,
North A.,
Webster T.,
HogenEsch H.,
Rundell A.
Publication year - 2003
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.10760
Subject(s) - succinimide , chemistry , antibody , antigen , chromatography , adsorption , combinatorial chemistry , biochemistry , biophysics , organic chemistry , immunology , biology
This study investigates the influence of antibody immobilization methods on antigen capture. Adsorption and two surface chemistries, an aminosilane chemistry and a common heterobifunctional crosslinker (N‐gamma‐maleimidobutyryloxy‐succinimide ester, GMBS), were compared and evaluated for their ability to immobilize antibodies and capture antigen. The role of protein A as an orienting protein scaffold component in each of these techniques was also evaluated. Through experimentation it was determined that the GMBS technique immobilized the highest amount of antibody and minimized nonspecific binding. For all techniques, the most functional antibodies were found to be those immobilized with protein A. Interestingly, the aminosilane technique demonstrated the highest antigen capture with antibody alone but also exhibited the highest level of nonspecific binding. © 2003 Wiley Periodicals. Biotechnol Bioeng 84: 215–223, 2003.