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Label-free and dynamic detection of biomolecular interactions for high-throughput microarray applications.

Proceedings of the National Academy of Sciences of the United States of America (2008-06-05)
Emre Ozkumur, James W Needham, David A Bergstein, Rodrigo Gonzalez, Mario Cabodi, Jonathan M Gershoni, Bennett B Goldberg, M Selim Unlü
RÉSUMÉ

Direct monitoring of primary molecular-binding interactions without the need for secondary reactants would markedly simplify and expand applications of high-throughput label-free detection methods. A simple interferometric technique is presented that monitors the optical phase difference resulting from accumulated biomolecular mass. As an example, 50 spots for each of four proteins consisting of BSA, human serum albumin, rabbit IgG, and protein G were dynamically monitored as they captured corresponding antibodies. Dynamic measurements were made at 26 pg/mm(2) SD per spot and with a detectable concentration of 19 ng/ml. The presented method is particularly relevant for protein microarray analysis because it is label-free, simple, sensitive, and easily scales to high-throughput.

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Albumine from human serum, lyophilized powder, ≥97% (agarose gel electrophoresis)