Surface-immobilized antibody-antigen binding studies by single molecule fluorescence imaging

Jamshid Temirov; Andrew Bradbury; James Werner

doi:10.1117/12.645334

27 February 2006 Surface-immobilized antibody-antigen binding studies by single molecule fluorescence imaging

Jamshid Temirov, Andrew Bradbury, James Werner

Proceedings Volume 6092, Ultrasensitive and Single-Molecule Detection Technologies; 60920O (2006) https://doi.org/10.1117/12.645334
Event: SPIE BiOS, 2006, San Jose, California, United States

Abstract

In this work we report Single Molecule (SM) microscopy studies of binding events between individual surface immobilized antibodies and fluorescent antigens using Total Internal Reflection (TIR) microscopy. Specific binding events of single biotin-conjugated 1quantum dots to anti-biotin antibodies, immobilized on an amine-coated cover glass using a heterobifunctional photo-reactive cross-linker were observed. The methodology of calculating the binding affinity (dissociation constant) from the time series of 2-D images is described.

Citation Download Citation

Jamshid Temirov, Andrew Bradbury, and James Werner "Surface-immobilized antibody-antigen binding studies by single molecule fluorescence imaging", Proc. SPIE 6092, Ultrasensitive and Single-Molecule Detection Technologies, 60920O (27 February 2006); https://doi.org/10.1117/12.645334

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