TY - JOUR
T1 - Oriented coupling of major histocompatibility complex (MHC) to sensor surfaces using light assisted immobilisation technology
AU - Snabe, Torben
AU - Røder, Gustav Andreas
AU - Neves-Petersen, Maria Teresa
AU - Buus, Søren
AU - Petersen, Steffen Bjørn
N1 - Keywords: Adsorption; Biosensing Techniques; Biotechnology; Coated Materials, Biocompatible; Crystallization; Histocompatibility Antigens Class I; Protein Array Analysis; Protein Binding; Ultraviolet Rays
PY - 2005
Y1 - 2005
N2 - Controlled and oriented immobilisation of proteins for biosensor purposes is of extreme interest since this provides more efficient sensors with a larger density of active binding sites per area compared to sensors produced by conventional immobilisation. In this paper oriented coupling of a major histocompatibility complex (MHC class I) to a sensor surface is presented. The coupling was performed using light assisted immobilisation--a novel immobilisation technology which allows specific opening of particular disulphide bridges in proteins which then is used for covalent bonding to thiol-derivatised surfaces via a new disulphide bond. Light assisted immobilisation specifically targets the disulphide bridge in the MHC-I molecule alpha(3)-domain which ensures oriented linking of the complex with the peptide binding site exposed away from the sensor surface. Structural analysis reveals that a similar procedure can be used for covalent immobilisation of MHC class II complexes. The results open for the development of efficient T cell sensors, sensors for recognition of peptides of pathogenic origin, as well as other applications that may benefit from oriented immobilisation of MHC proteins.
AB - Controlled and oriented immobilisation of proteins for biosensor purposes is of extreme interest since this provides more efficient sensors with a larger density of active binding sites per area compared to sensors produced by conventional immobilisation. In this paper oriented coupling of a major histocompatibility complex (MHC class I) to a sensor surface is presented. The coupling was performed using light assisted immobilisation--a novel immobilisation technology which allows specific opening of particular disulphide bridges in proteins which then is used for covalent bonding to thiol-derivatised surfaces via a new disulphide bond. Light assisted immobilisation specifically targets the disulphide bridge in the MHC-I molecule alpha(3)-domain which ensures oriented linking of the complex with the peptide binding site exposed away from the sensor surface. Structural analysis reveals that a similar procedure can be used for covalent immobilisation of MHC class II complexes. The results open for the development of efficient T cell sensors, sensors for recognition of peptides of pathogenic origin, as well as other applications that may benefit from oriented immobilisation of MHC proteins.
U2 - 10.1016/j.bios.2005.06.010
DO - 10.1016/j.bios.2005.06.010
M3 - Journal article
C2 - 16139496
SN - 0956-5663
VL - 21
SP - 1553
EP - 1559
JO - Biosensors and Bioelectronics
JF - Biosensors and Bioelectronics
IS - 8
ER -