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

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.