Show simple item record

dc.contributor.authorRead, Thomas
dc.contributor.authorOlkhov, RV
dc.contributor.authorWilliamson, E. Diane
dc.contributor.authorShaw, AM
dc.date.accessioned2015-08-18T15:38:51Z
dc.date.issued2015-07-19
dc.description.abstractA unified approach to affinity screening for Fab and Fc interactions of an antibody for its antigen and FcγR receptor has been developed. An antigen array is used for the Fab affinity and cross-reactivity screening and protein A/G proxy is the FcγR receptor. The affinities are derived using a simple 1:1 binding model with a consistent error analysis. The association and dissociation kinetics are measured over optimised times for accurate determination. The Fab/Fc affinities are derived for ten antibodies: mAb-actin (mouse), pAb-BSA (sheep), pAb-collagen V (rabbit), pAb-CRP (goat), mAb-F1 (mouse), mAbs (mouse) 7.3, 12.3, 29.3, 36.3 and 46.3 raised against LcrV in Yersinia pestis. The rate of the dissociation of antigen–antibody complexes relates directly to their immunological function as does the Fc-FcγR complex and a new half-life plot has been defined with a Fab/Fc half-life range of 17–470 min. The upper half-life value points to surface avidity. Two antibodies that are protective as an immunotherapy define a Fab half-life >250 min and an Fc half-life >50 min as characteristics of ideal interactions which can form the basis of an antibody screen for immunotherapy.en_GB
dc.description.sponsorshipBiotechnology and Biological Sciences Research Council (BBSRC)en_GB
dc.identifier.citationVol. 407 (24), pp 7349-7357
dc.identifier.doi10.1007/s00216-015-8897-6
dc.identifier.urihttp://hdl.handle.net/10871/18067
dc.language.isoenen_GB
dc.publisherSpringer Verlagen_GB
dc.rights.embargoreasonPublisher policyen_GB
dc.rightsCopyright © Springer-Verlag Berlin Heidelberg 2015en_GB
dc.subjectBioanalytical methodsen_GB
dc.subjectImmunoassays/ELISAen_GB
dc.subjectOptical sensorsen_GB
dc.titleLabel-free Fab and Fc affinity/avidity profiling of the antibody complex half-life for polyclonal and monoclonal efficacy screeningen_GB
dc.typeArticleen_GB
dc.identifier.issn1618-2642
dc.descriptionThe final publication is available at Springer via http://dx.doi.org/10.1007/s00216-015-8897-6en_GB
dc.identifier.eissn1618-2650
dc.identifier.journalAnalytical and Bioanalytical Chemistryen_GB


Files in this item

This item appears in the following Collection(s)

Show simple item record