Automated maskless photolithography system for peptide microarray synthesis on a chip.
Maskless photolithographic peptide synthesis was performed on a glass chip using an automated peptide array synthesizer system. The peptide array synthesizer was built in a closed box, which contained optical and fluidic systems. The conditions for peptide synthesis were fully controlled by a computer program. For the peptide synthesis on a glass chip, 20 NVOC-protected amino acids were synthesized. The coupling efficiencies of two model peptide sequences were examined on ACA/APTS and PEG/CHI/GPTS chips. PEG/CHI/GPTS chip gave higher average stepwise yields of GIYWHHY (94%) and YIYGSFK (98%) than those of ACA/APTS chip. To quantify peptide-protein binding affinity, HPQ- or HPM-containing pentapeptides were synthesized on a PEG/CHI/GPTS chip and the binding event of Cy3 labeled-streptavidin was quantified. The peptide sequence of IQHPQ showed highest binding affinity with Cy3 labeled-streptavidin. The results demonstrated that the photolithographic peptide array synthesis method efficiently quantified the binding activities of protein-peptide interactions and it can be used for additional biological assay applications.
Shin
Dong-Sik
DS
School of Chemical and Biological Engineering and School of Electrical Engineering and Computer Science, Seoul National University, Seoul 151-744, Korea.
Lee
Kook-Nyung
KN
Yoo
Byung-Wook
BW
Kim
Jaehi
J
Kim
Mira
M
Kim
Yong-Kweon
YK
Lee
Yoon-Sik
YS
D015394:Molecular Structure; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry; D055668:Photochemical Processes; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods*
D010455:Peptides