The structural orientation of antibody layers bound to engineered biosensor surfaces

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dc.contributor.authorLe Brun, APen_AU
dc.contributor.authorHolt, SAen_AU
dc.contributor.authorShah, DSHen_AU
dc.contributor.authorMajkrzak, CFen_AU
dc.contributor.authorLakey, JHen_AU
dc.date.accessioned2011-09-21T04:54:29Zen_AU
dc.date.available2011-09-21T04:54:29Zen_AU
dc.date.issued2011-04-01en_AU
dc.date.statistics2011-09-21en_AU
dc.description.abstractThis paper describes a membrane protein array that binds immunoglobulin G at its constant regions whilst leaving the variable regions free to bind antigen. The scaffold of the array is the transmembrane domain of outer membrane protein A (tOmpA) from Escherichia colt engineered to assemble as an oriented monolayer on gold surfaces via a single cysteine residue. Other protein domains can be fused to the N and C termini of the scaffold. In this study we use circularly permuted ctOmpA fused to two Z domains of Staphylococcus aureus protein A (ZZctOmpA) to create the immunoglobulin G-binding array. The solution structure of the engineered proteins was assessed by circular dichroism spectroscopy. Assembly of the array, attachment of antibodies and antigen binding were measured using surface plasmon resonance and neutron reflection. Compared to mouse IgG2, polyclonal IgG from rabbit bound very strongly to ZZctOmpA and the dissociation of the immunoglobulin was slow enough to allow neutron reflection studies of the assembled layer with antigen. Using both magnetic and isotopic contrasts a complete layer by layer model was defined which revealed that the 223 A high layer contains antibodies in an upright orientation. (C) 2011 Elsevier Ltd. All rights reserved.en_AU
dc.identifier.citationLe Brun, A.P., Holt, S.A., Shah, D.S.H., Majkrzak, C.F., Lakey, J.H. (2011). The structural orientation of antibody layers bound to engineered biosensor surfaces, Biomaterials, 32(12), 3303-3311. doi:10.1016/j.biomaterials.2011.01.026en_AU
dc.identifier.govdoc3415en_AU
dc.identifier.issn0142-9612en_AU
dc.identifier.issue12en_AU
dc.identifier.journaltitleBiomaterialsen_AU
dc.identifier.pagination3303-3311en_AU
dc.identifier.urihttp://dx.doi.org/10.1016/j.biomaterials.2011.01.026en_AU
dc.identifier.urihttp://apo.ansto.gov.au/dspace/handle/10238/3763en_AU
dc.identifier.volume32en_AU
dc.language.isoenen_AU
dc.publisherElsevieren_AU
dc.subjectMembrane proteinsen_AU
dc.subjectSensorsen_AU
dc.subjectSpectroscopyen_AU
dc.subjectImmunoglobulinsen_AU
dc.subjectPlasmonsen_AU
dc.subjectAntibodiesen_AU
dc.titleThe structural orientation of antibody layers bound to engineered biosensor surfacesen_AU
dc.typeJournal Articleen_AU
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