Oriented coupling of major histocompatibility complex (MHC) to sensor surfaces using light assisted immobilisation technology.
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.
Snabe
Torben
T
Aalborg University, Institute of Physics and Nanotechnology, Biostructure and Protein Engineering, Denmark.
Røder
Gustav Andreas
GA
Neves-Petersen
Maria Teresa
MT
Buus
Søren
S
Petersen
Steffen Bjørn
SB
D000327:Adsorption / Q000528:radiation effects; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D001709:Biotechnology / Q000295:instrumentation / Q000379:methods*; D020099:Coated Materials, Biocompatible / Q000737:chemistry* / Q000528:radiation effects; D003460:Crystallization / Q000379:methods*; D015395:Histocompatibility Antigens Class I / Q000032:analysis* / Q000737:chemistry* / Q000528:radiation effects; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods*; D011485:Protein Binding / Q000187:drug effects; D014466:Ultraviolet Rays
D020099:Coated Materials, Biocompatible; D015395:Histocompatibility Antigens Class I