Biocompatibility of surfaces for antibody microarrays: design of macroporous silicon substrates.
Major efforts to develop antibody microarray technology to enable global proteome analysis to be performed in a facile manner are under way. In this process, the design and the properties of the substrate will play crucial roles. In the present study, we have developed novel, highly biocompatible solid supports for microarrays, using adsorbed recombinant human single-framework antibody fragments as probes. Several silicon-based supports, including planar silicon, micro- and macroporous silicon, and nitrocellulose-coated variants thereof, were designed and evaluated in a stepwise procedure. The surfaces were scored based on biocompatibility and probe binding capacity as judged by spot morphology, signal intensities, signal to noise ratios, dynamic range, sensitivity, and reproducibility. A set of five commercially available substrates, selected to represent a set of supports providing different surface and coupling chemistries, was used as reference surfaces. The results showed that several well-performing silicon-based supports could be designed; in particular, a nitrocellulose-coated macroporous variant, MAP3-NC7, received the highest scores. In comparison, MAP3-NC7 displayed properties equal to or better than those of the reference substrates. Taken together, designed surfaces based on silicon can undoubtedly meet the requirements of the next generation of solid supports for antibody microarrays.
Steinhauer
Cornelia
C
Department of Immunotechnology, Lund University, P.O. Box 7031, SE-220 07 Lund, Sweden.
Ressine
Anton
A
Marko-Varga
György
G
Laurell
Thomas
T
Borrebaeck
Carl A K
CA
Wingren
Christer
C
D000906:Antibodies; D003101:Collodion / Q000737:chemistry*; D006801:Humans; D017646:Organosilicon Compounds / Q000737:chemistry*; D040081:Protein Array Analysis / Q000379:methods*; D040901:Proteomics / Q000379:methods*; D012825:Silicon / Q000737:chemistry*
D000906:Antibodies; D017646:Organosilicon Compounds; D003101:Collodion; D012825:Silicon