%0 Journal Article %1 Wassaf2006241 %A Wassaf, Dina %A Kuang, Guannan %A Kopacz, Kris %A Wu, Qi-Long %A Nguyen, Que %A Toews, Mark %A Cosic, Janja %A Jacques, Judith %A Wiltshire, Steve %A Lambert, Jeremy %A Pazmany, Csaba C. %A Hogan, Shannon %A Ladner, Robert C. %A Nixon, Andrew E. %A Sexton, Daniel J. %D 2006 %J Analytical Biochemistry %K Antibody display phage sprimimbpaper %N 2 %P 241--253 %R DOI: 10.1016/j.ab.2006.01.043 %T High-throughput affinity ranking of antibodies using surface plasmon resonance microarrays %U http://www.sciencedirect.com/science/article/B6W9V-4J7H34M-2/2/e88057dbddc99dfa8734f70244467a69 %V 351 %X A method was developed to rapidly identify high-affinity human antibodies from phage display library selection outputs. It combines high-throughput Fab fragment expression and purification with surface plasmon resonance (SPR) microarrays to determine kinetic constants (kon and koff) for 96 different Fab fragments in a single experiment. Fabs against human tissue kallikrein 1 (hK1, KLK1 gene product) were discovered by phage display, expressed in Escherichia coli in batches of 96, and purified using protein A PhyTip columns. Kinetic constants were obtained for 191 unique anti-hK1 Fabs using the Flexchip SPR microarray device. The highest affinity Fabs discovered had dissociation constants of less than 1 nM. The described SPR method was also used to categorize Fabs according to their ability to recognize an apparent active site epitope. The ability to rapidly determine the affinities of hundreds of antibodies significantly accelerates the discovery of high-affinity antibody leads.

@article{Wassaf2006241, abstract = {A method was developed to rapidly identify high-affinity human antibodies from phage display library selection outputs. It combines high-throughput Fab fragment expression and purification with surface plasmon resonance (SPR) microarrays to determine kinetic constants (kon and koff) for 96 different Fab fragments in a single experiment. Fabs against human tissue kallikrein 1 (hK1, KLK1 gene product) were discovered by phage display, expressed in Escherichia coli in batches of 96, and purified using protein A PhyTip columns. Kinetic constants were obtained for 191 unique anti-hK1 Fabs using the Flexchip SPR microarray device. The highest affinity Fabs discovered had dissociation constants of less than 1 nM. The described SPR method was also used to categorize Fabs according to their ability to recognize an apparent active site epitope. The ability to rapidly determine the affinities of hundreds of antibodies significantly accelerates the discovery of high-affinity antibody leads.}, added-at = {2009-09-01T05:30:05.000+0200}, author = {Wassaf, Dina and Kuang, Guannan and Kopacz, Kris and Wu, Qi-Long and Nguyen, Que and Toews, Mark and Cosic, Janja and Jacques, Judith and Wiltshire, Steve and Lambert, Jeremy and Pazmany, Csaba C. and Hogan, Shannon and Ladner, Robert C. and Nixon, Andrew E. and Sexton, Daniel J.}, biburl = {https://www.bibsonomy.org/bibtex/23a1fe2be74ac65336aa2120c0923c03f/clausted}, description = {ScienceDirect - Analytical Biochemistry : High-throughput affinity ranking of antibodies using surface plasmon resonance microarrays}, doi = {DOI: 10.1016/j.ab.2006.01.043}, interhash = {5eb9ff35e3c9ad96b1a31b79eae01cdc}, intrahash = {3a1fe2be74ac65336aa2120c0923c03f}, issn = {0003-2697}, journal = {Analytical Biochemistry}, keywords = {Antibody display phage sprimimbpaper}, number = 2, pages = {241--253}, timestamp = {2009-09-01T05:30:05.000+0200}, title = {High-throughput affinity ranking of antibodies using surface plasmon resonance microarrays}, url = {http://www.sciencedirect.com/science/article/B6W9V-4J7H34M-2/2/e88057dbddc99dfa8734f70244467a69}, volume = 351, year = 2006 }