Double chip protein arrays using recombinant single-chain Fv antibody fragments.
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.
Gilbert
Ilka
I
nanotype, Graefelfing, Germany.
Schiffmann
Susanne
S
Rubenwolf
Susanne
S
Jensen
Kristian
K
Mai
Thao
T
Albrecht
Christian
C
Lankenau
Andreas
A
Beste
Gerald
G
Blank
Kerstin
K
Gaub
Hermann E
HE
Clausen-Schaumann
Hauke
H
D019921:Biotinylation; D007128:Immunoglobulin Fragments / Q000276:immunology*; D007135:Immunoglobulin Variable Region / Q000235:genetics* / Q000276:immunology; D040081:Protein Array Analysis / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D012680:Sensitivity and Specificity
D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins; C055966:immunoglobulin Fv