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Immobilization of Proteins for Biosensor Development

  • Chapter
Immobilized Biosystems

Abstract

There is considerable interest in the permanent attachment of bio-molecules to solid surfaces because of the many uses of immobilized proteins: for purification of molecules that bind to antibodies, lectins or other binding proteins; in solid-phase analytical assays; for improved biocompatibility of materials; and to provide selectivity to biosensors. In such applications, the amount of protein immobilized and the retention of physiological activity of the immobilized protein (such as binding of a second molecule) are of interest. This work will report the amount and activity of immobilized proteins as a function of the method of immobilization, using instrumental analytical techniques. The results are of interest to all whose work involves immobilized proteins, but the study was inspired by its application to the field of biosensors, as evidenced by the choice of solid surfaces (quartz and silicon), and the types of proteins employed (those which bind a second molecule).

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Abbreviations

ab:

antibody (anti-rabbit immunoglobulin G (whole molecule) FITC conjugate, developed from goat)

ag:

antigen (rabbit immunoglobulin G)

APTES:

3-aminopropyl triethoxy silane

BSA:

bovine serum albumin

Con A:

concanavalin A

FITC:

fluorescein isothiocyanate

glut:

glutaraldehyde-derivatized surface

glutaralde:

glutaraldehyde-derivatized surface

HABA:

2-[4-hydroxyphenylazo]benzoic acid

IgG:

immunoglobulin G

ligand:

a molecule bound by a receptor

receptor:

includes molecular receptors, antibodies, enzymes and lectins

RT-APTES:

APTES deposited under room temperature conditions

TEA-APTES:

APTES deposited under reflux in the presence of the base catalyst triethylamine

tresyl:

2,2,2-trifluoroethane sulfonyl

tresyl:

chloride 2,2,2-trifluoroethane sulfonyl chloride

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Authors and Affiliations

  1. Chemical Sensors Group, Department of Chemistry, University of Toronto, Mississauga, Canada

    Elaine T. Vandenberg, R. Stephen Brown & Ulrich J. Krull

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  1. Elaine T. Vandenberg
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  2. R. Stephen Brown
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  1. Tivelco International Inc., Ottawa, Canada

    Ivan A. Veliky

  2. Department of Biology, Southwest Texas State University, San Marcos, Texas, USA

    Robert J. C. McLean

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Vandenberg, E.T., Brown, R.S., Krull, U.J. (1994). Immobilization of Proteins for Biosensor Development. In: Veliky, I.A., McLean, R.J.C. (eds) Immobilized Biosystems. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-1334-2_2

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