Spot arrays on modified glass surfaces for efficient SPOT synthesis and on-chip bioassay of peptides.
To make SPOT synthesis of peptides and their assays on glass surfaces more convenient, a simple method for making spot arrays on a slide glass was designed through patterning with a photoresist and perfluorination followed by amination with various silane compounds and polymers. With these spot-arrayed glass surfaces, we could measure the coupling completion of each Fmoc amino acid on the glass surface by direct fluorescence analysis after fluorescence-labeling the amino groups on the surface of each spot. Then we synthesized several types of decapeptides and HPQ-pentapeptides on the spot-arrayed glasses and identified the optimal surface condition for stepwise peptide coupling and on-chip bioassay. After optimizing the surface conditions, we synthesized a model library of biotin-Gly-Ala-P(1)-Gly (P(1): one of 19 amino acids) and successfully replicated the well-known alpha-chymotrypsin subsite specificities through Cy5-streptavidin binding to the remaining biotin on the surface after the enzymatic digestion.
Kim
Do-Hyun
DH
School of Chemical and Biological Engineering, Seoul National University, Seoul 151-744, Republic of Korea.
Shin
Dong-Sik
DS
Lee
Yoon-Sik
YS
D000596:Amino Acids / Q000737:chemistry; D019921:Biotinylation; D048271:Chitosan / Q000737:chemistry; D002918:Chymotrypsin / Q000378:metabolism; D020650:Combinatorial Chemistry Techniques; D005449:Fluorenes / Q000737:chemistry; D005898:Glass; D010455:Peptides / Q000138:chemical synthesis*; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods*; D012821:Silanes; D019809:Streptavidin / Q000737:chemistry
C403136:3-glycidoxypropyltrimethoxysilane; D000596:Amino Acids; D005449:Fluorenes; C016456:N(alpha)-fluorenylmethyloxycarbonylamino acids; D010455:Peptides; D012821:Silanes; D048271:Chitosan; D019809:Streptavidin; D002918:Chymotrypsin; C117608:alpha-chymotrypsin