Share Email Print

Proceedings Paper

Waveguide multichannel immunoassay using photo-deprotection immobilization
Author(s): Thomas E. Plowman; Amy S. Blawas; Tom F. Oliver; W. Monty Reichert
Format Member Price Non-Member Price
PDF $17.00 $21.00

Paper Abstract

A planar optical waveguide was used to simultaneously excite fluorescence due to antigen binding in three separate areas of immobilized antibody. Biotin labeled, polyclonal antibodies to goat, human, and rabbit IgG were immobilized through surface bound, photo-activated MeNPOC-biotin-bSA and avidin. Exposing the MeNPOC to UV light effectively uncaged the biotin molecule attached to the bSA and allowed avidin, followed by the biotin labeled antibody, to bind to the waveguide surface. Whereas a time intensive, non-specific binding prone step-and-repeat method is normally used to form the individual capture layers, we chose to pursue a combined deposition method involving sample wells and photo- activated crosslinkers. The result was a covalently linked multi-component capture layer formed in a short period of the time. Specific and cross-reactive activities of this antibody array were gauged by sequentially injecting analyte specific to one antibody area at a time. Results suggested that the binding of each analyte occurred predominately in the correct area and, depending on the particular antibody, generated varying levels of cross reactivity. A comparison of result with previously acquired, physically adsorbed capture layer data did not infer one deposition technique was better than the other.

Paper Details

Date Published: 21 April 1999
PDF: 7 pages
Proc. SPIE 3603, Systems and Technologies for Clinical Diagnostics and Drug Discovery II, (21 April 1999); doi: 10.1117/12.346737
Show Author Affiliations
Thomas E. Plowman, Duke Univ. (United States)
Amy S. Blawas, Duke Univ. (United States)
Tom F. Oliver, Duke Univ. (United States)
W. Monty Reichert, Duke Univ. (United States)

Published in SPIE Proceedings Vol. 3603:
Systems and Technologies for Clinical Diagnostics and Drug Discovery II
Gerald E. Cohn; John C. Owicki, Editor(s)

© SPIE. Terms of Use
Back to Top