Generating recombinant anti-idiotypic antibodies for the detection of haptens in solution.
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.
Raats
Jos
J
Department of Biochemistry, University of Nijmegen, Nijmegen, The Netherlands. j.raats@ncmls.kun.nl
van Bree
Nicole
N
van Woezik
Jochem
J
Pruijn
Ger
G
D000450:Aldosterone / Q000097:blood / Q000276:immunology; D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D000918:Antibody Specificity; D001667:Binding, Competitive; D002645:Chickens; D000431:Ethanol / Q000494:pharmacology; D006241:Haptens / Q000032:analysis* / Q000097:blood / Q000276:immunology; D006801:Humans; D006845:Hydrocarbons, Fluorinated / Q000494:pharmacology; D006854:Hydrocortisone / Q000097:blood / Q000276:immunology; D007118:Immunoassay / Q000379:methods; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D007074:Immunoglobulin G / Q000235:genetics / Q000276:immunology; D009841:Oligonucleotides / Q000235:genetics; D019151:Peptide Library; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D013739:Testosterone / Q000378:metabolism; D014357:Trypsin / Q000235:genetics / Q000378:metabolism
D000888:Antibodies, Anti-Idiotypic; D006241:Haptens; D006845:Hydrocarbons, Fluorinated; D007128:Immunoglobulin Fragments; D007074:Immunoglobulin G; D009841:Oligonucleotides; D019151:Peptide Library; D011994:Recombinant Proteins; D000431:Ethanol; D013739:Testosterone; D000450:Aldosterone; D014357:Trypsin; D006854:Hydrocortisone