Detection of porphyrin using a short peptide immobilized on a surface plasmon resonance sensor chip.
In this paper the development and feasibility of a novel detection system for a low molecular weight chemical, in which a peptide was utilized as a binding molecule, are described. Surface plasmon resonance (SPR) apparatus was used as a transducer. The porphyrin binding peptide, PSP2, was used as a model peptide ligand, while a porphyrin derivative, H(2)TMpyP, was used as a model low-molecular-weight chemical. PSP2 was covalently immobilized onto the SPR sensor chip and SPR measurement using the PSP2-immobilized chip for various concentrations of porphyrin was carried out. H(2)TMpyP was detectable in the range from 100 ng ml(-1) to 10 microg ml(-1) with a linear correlation and good precision and the PSP2-immobilized chip could be regenerated within 1 min after measurement in this system. From comparison of the detection manners of three porphyrin derivatives, the ability of a short peptide to discriminate between differences in molecular structure was demonstrated. Moreover, the self-assembled monolayer (SAM) of PSP2 was successfully prepared on the gold substrate and H(2)TMpyP could be detected using the PSP2-SAM chip.
Nakamura
C
C
National Institute for Advanced Interdisciplinary Research, AIST, MITI, 1-1-4 Higashi, Tsukuba 305-8562, Japan. chikashi_nakamura@aist.go.jp
Inuyama
Y
Y
Shirai
K
K
Sugimoto
N
N
Miyake
J
J
D000595:Amino Acid Sequence; D002352:Carrier Proteins / Q000737:chemistry; D008024:Ligands; D015394:Molecular Structure; D009842:Oligopeptides / Q000737:chemistry; D011166:Porphyrins / Q000032:analysis* / Q000737:chemistry; D011524:Protoporphyrins / Q000032:analysis / Q000737:chemistry; D020349:Surface Plasmon Resonance / Q000379:methods*
D002352:Carrier Proteins; D008024:Ligands; D009842:Oligopeptides; D011166:Porphyrins; D011524:Protoporphyrins; C021096:tetra(4-N-methylpyridyl)porphine; C029972:alpha,beta,gamma,delta-tetrakis(4-N-trimethylaminophenyl)porphine; C028025:protoporphyrin IX