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

Abstract

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 prescreened 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.