title: Oriented coupling of major histocompatibility complex (MHC) to sensor surfaces using light assisted immobilisation technology.
abstract: 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.
authors.affiliation: Aalborg University, Institute of Physics and Nanotechnology, Biostructure and Protein Engineering, Denmark.
authors.forename: Torben
authors.initials: T
authors.lastname: Snabe
authors.forename: Gustav Andreas
authors.initials: GA
authors.lastname: Røder
authors.forename: Maria Teresa
authors.initials: MT
authors.lastname: Neves-Petersen
authors.forename: Søren
authors.initials: S
authors.lastname: Buus
authors.forename: Steffen Bjørn
authors.initials: SB
authors.lastname: Petersen
mesh_terms: 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
chemical_list: D020099:Coated Materials, Biocompatible; D015395:Histocompatibility Antigens Class I