A new polymeric coating for protein microarrays.
Despite the increasing interest in arraying proteins in a high-density format, several technical issues still impede the development of protein microarray technology. One of the major problems is the availability of substrates that are able to bind native proteins with high density. In this study, we investigated the suitability of a novel surface as a support for protein microarrays. A polymeric glass coating is obtained by physical adsorption of a N,N-dimethylacrylamide (DMA), N,N-acryloyloxysuccinimide (NAS), and [3-(methacryloyl-oxy)propyl]trimethoxysilyl (MAPS) copolymer. The coating procedure provides a fast and inexpensive method of producing hydrophilic functional surfaces. The slide performance was investigated in a protein-protein interaction experiment and in the assessment of rheumatoid factor (RF) in human serum samples. The results demonstrate that the ligands immobilized on the polymeric surface maintain an active conformation and are easily accessible, providing a detection limit of 54amol/spot. Moreover, in the RF assay, after hybridization with the sera, the slides have a low background, leading to a detection limit of 900amol/spot.
Cretich
Marina
M
Istituto di Chimica del Riconoscimento Molecolare (ICRM), C.N.R., Via Mario Bianco 9, 20131 Milano, Italy. marina.cretich@icrm.cnr.it
Pirri
Giovanna
G
Damin
Francesco
F
Solinas
Isabella
I
Chiari
Marcella
M
D000178:Acrylamides; D007118:Immunoassay / Q000379:methods; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D011108:Polymers; D040081:Protein Array Analysis / Q000295:instrumentation* / Q000379:methods
D000178:Acrylamides; D007140:Immunoglobulin Fab Fragments; D011108:Polymers; C099046:N,N-dimethylacrylamide