Portal de Periódicos da CAPES

Generating Recombinant Anti-idiotypic Antibodies for the Detection of Haptens in Solution

2003; Taylor & Francis; Volume: 24; Issue: 2 Linguagem: Inglês

10.1081/ias-120020081

1532-4230

Jos Raats, Nicole van Bree, Jochem van Woezik, Ger J.M. Pruijn,

Glycosylation and Glycoproteins Research

Abstract 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. Keywords: Phage displayHaptensCortisolAldosteroneCompetition assay Acknowledgments We would like to thank Prof. Winter and Dr. Nissim for the use of the synthetic library. We thank Dr. Kristensen for the trypsin-sensitive helper phage. We thank the CSHL phage display course team (Carlos Barbas, Don Siegel, and Gregg Silverman) for providing us with both the knowledge for the construction of the chicken phage display libraries as well as for the pComb3H vector system for cloning the chicken libraries. We thank Future Diagnostics B.V., Wijchen, The Netherlands, for financial support. This work was partially funded by BTS (Bedrijfsgerichte Technologische Samenwerking) grant 97247, and the Royal Netherlands Academy of Arts and Sciences (KNAW).