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Generating recombinant anti-idiotypic antibodies for the detection of haptens in solution.

Posted by on in 2003
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Raats J, van Bree N, van Woezik J, Pruijn G J Immunoassay Immunochem 2003 24:115-46

Abstract

A new method is described for generating recombinant human and chicken antibody fragments for accurate quantification of haptens in solution. The chemistry of labelling small molecules has always been a problem in the development of immunoassays. Here, we describe a specific panning procedure that enables the selection of recombinant anti-idiotypic phage antibodies that bind to hapten binding molecules (e.g., antibodies) in the absence of the hapten, but are displaced in a highly specific and concentration dependent manner, in the presence of the hapten. The major advantage of such a detection system is that there is no need to label the hapten or to covalently attach it to a solid phase. In this study we demonstrate, using cortisol and aldosterone as model haptens, that the recombinant antibody phage display technology offers great possibilities to generate recombinant anti-idiotypic antibodies. Furthermore, we show that such antibodies can be used successfully to design highly sensitive immunoassays for the quantification of small molecules.

Author Address

Department of Biochemistry, University of Nijmegen, Nijmegen, The Netherlands. j.raats@ncmls.kun.nl.

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