The application of neoglycopeptides in the development of sensitive surface plasmon resonance-based biosensors.
The development of a biosensor based on surface plasmon resonance is described for the detection of carbohydrate-binding proteins in solution on a Biacore 2000 instrument, using immobilized glycopeptides as ligands. Their selection was based on previous screenings of solid-phase glycopeptide libraries with Ricinus communis agglutinin (RCA(120)) and human adhesion/growth-regulatory galectin-1 (h-Gal-1). Glycopeptides were immobilized on Au sensor chips functionalized with mixed self-assembled monolayers of different ratios of 11-mercapto-1-undecanol and 11-mercaptoundecanoic acid, and of 3-mercapto-1-propanol and 11-mercaptoundecanoic acid. The biosensors were optimized for the detection of RCA(120), and a detection limit of 0.13 nM was obtained. Subsequent experiments with h-Gal-1 indicated a detection limit of at least 0.9 nM for this lectin. Additionally, the effect of interfering proteins on the sensitivity of the optimized biosensor was investigated.
Maljaars
C Elizabeth P
CE
Bijvoet Center, Department of Bio-Organic Chemistry, Utrecht University, Utrecht, The Netherlands.
de Souza
Adriana Carvalho
AC
Halkes
Koen M
KM
Upton
Peter J
PJ
Reeman
Stuart M
SM
André
Sabine
S
Gabius
Hans-Joachim
HJ
McDonnell
Martin B
MB
Kamerling
Johannis P
JP
D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D002352:Carrier Proteins / Q000032:analysis; D037483:Galectin 1 / Q000032:analysis; D006020:Glycopeptides / Q000737:chemistry*; D037121:Plant Lectins / Q000032:analysis; D012680:Sensitivity and Specificity; D020349:Surface Plasmon Resonance / Q000379:methods*
D002352:Carrier Proteins; D037483:Galectin 1; D006020:Glycopeptides; D037121:Plant Lectins; C017707:Ricinus communis agglutinin-1