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Double chip protein arrays using recombinant single-chain Fv antibody fragments

: Gilbert, I.; Schiffmann, S.; Rubenwolf, S.; Jensen, K.; Mai, T.; Albrecht, C.; Lankenau, A.; Beste, G.; Blank, K.; Gaub, H.E.; Clausen-Schaumann, H.


Görg, A.:
Proteomic Forum 2003, Proceedings of the International Meeting on Proteome Analysis : Munich, Germany, 14 - 17 September 2003
Weinheim: Wiley-VCH, 2004 (Proteomics 4.2004, Nr.5)
International Meeting on Proteome Analysis <3, 2003, München>
Conference Paper, Journal Article
Fraunhofer IBMT ()

Protein arrays permit the parallel analysis of many different markers in a small sample volume. However, the problem of cross-reactivity limits the degree of multiplexing in parallel sandwich immunoassays (using monoclonal antibodies (mAbs)), meaning antibodies must be pre-screened in order to reduce false positives. In contrast, we use a second chip surface for the local application of detection antibodies, thereby efficiently eliminating antibody cross-reactions. Here, we illustrate the potential advantages of using single-chain Fv fragments rather than mAbs as capture and detection molecules with this double chip technology.