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Proceedings Paper

Specific immobilization of human immunoglobulin G on gold-coated silicon microcantilever array
Author(s): Sandeep Kumar Vashist; Rupinder Tewari; Ram Prakash Bajpai; Lalit Mohan Bharadwaj; Roberto Raiteri
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Paper Abstract

We demonstrate a procedure for immobilizing human immunoglobulin G (IgG) on an array of gold-coated silicon microcantilevers. The procedure employed protein A for the specific immobilization of human IgG on the gold surface. Protein A bound specifically to the gold-coated upper surface of the silicon microcantilever and had no interaction with the silicon surface. It binds to the constant Fc regions of human IgG keeping the antigen binding sites on the variable Fab region free to bind to antigens. Fluorescent microscopy was done to analyze qualitatively the biomolecular binding of human IgG using FITC labeled goat anti-human IgG. The immobilization densities of protein A and human IgG were 112±19 ng/cm2 and 629±23ng/cm2, as determined employing horse radish peroxidase (HRP) labeled biomolecules by 3, 3', 4, 4'-tetramethyl benzidine (TMB) substrate assay. The uniformness of the biomolecular coatings was further determined by atomic force microscopy (AFM). Surface plasmon resonance (SPR) was used to cross-validate the immobilization density of functional human IgG molecules immobilized on the gold surface w.r.t. that obtained by TMB substrate assay.

Paper Details

Date Published: 22 January 2007
PDF: 7 pages
Proc. SPIE 6465, Microfluidics, BioMEMS, and Medical Microsystems V, 64650G (22 January 2007); doi: 10.1117/12.695439
Show Author Affiliations
Sandeep Kumar Vashist, Univ. of Genoa (Italy)
Rupinder Tewari, Panjab Univ. (India)
Ram Prakash Bajpai, Central Scientific Instruments Organisation (India)
Lalit Mohan Bharadwaj, Central Scientific Instruments Organisation (India)
Roberto Raiteri, Univ. of Genoa (Italy)

Published in SPIE Proceedings Vol. 6465:
Microfluidics, BioMEMS, and Medical Microsystems V
Ian Papautsky; Wanjun Wang, Editor(s)

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