Production of a single-chain variable fragment antibody against fumonisin B1.
The selection of synthetic antibody fragments from large phage libraries has become a common method for the generation of specific antibodies. The technique is particularly valuable when antibodies against small, non-immunogenic molecules (haptens) or highly toxic substances have to be produced. In addition, haptens are usually coupled to protein carriers, bearing the risk that the free hapten is not detectable. Here, a single variable chain antibody (scFv) against the highly toxic mycotoxin fumonisin B1 has been produced. The hapten was coupled via a linker to biotin. Using this conjugate and a naive scFv library, it was possible to circumvent both the necessity of immunization and the risk of a disguised hapten. The scFv obtained after three panning rounds was found to bind specifically to both free fumonisin B1 and fumonisin-biotin conjugate. Also fumonisin B2 was bound by the scFv. Modeling of both scFv and fumonisin B1 molecule revealed a good fitting of structures. The antibody obtained can potentially be used for developing a rapid and affordable immunoassay for detection of food contamination and can be applied in immunoaffinity chromatography, usually carried out prior to HPLC analysis of mycotoxin-contaminated food and feed.
Lauer
Björn
B
LG Molekulargenetik, University of Hannover, Herrenhäuser Strasse 2, D-30419 Hannover, Germany.
Ottleben
Ilka
I
Jacobsen
Hans-Jörg
HJ
Reinard
Thomas
T
D001710:Biotin / Q000276:immunology; D037341:Fumonisins / Q000032:analysis / Q000737:chemistry / Q000276:immunology*; D006241:Haptens / Q000276:immunology; D007128:Immunoglobulin Fragments / Q000096:biosynthesis* / Q000737:chemistry; D007135:Immunoglobulin Variable Region / Q000276:immunology*; D008958:Models, Molecular; D019151:Peptide Library
D037341:Fumonisins; D006241:Haptens; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; C056933:fumonisin B1; D001710:Biotin