Influence of format on in vitro penetration of antibody fragments through porcine cornea

dc.contributor.authorBrereton, Helen Mary
dc.contributor.authorTaylor, Susan D
dc.contributor.authorFarrall, Alexandra L
dc.contributor.authorHocking, Dianna M
dc.contributor.authorThiel, Michael A
dc.contributor.authorTea, Melinda Nay
dc.contributor.authorCoster, Douglas John
dc.contributor.authorWilliams, Keryn Anne
dc.description.abstractAIM: Antibody fragments, appropriately formulated, can penetrate through the ocular surface and thus have potential as therapeutic agents. The aim was to investigate the influence of protein fragment format on the kinetics and extent of ocular penetration in vitro. METHODS: Immunoglobulin single chain variable domain fragments of a murine monoclonal antibody with specificity for rat CD4 were engineered with a 20 or 11 amino acid linker by assembly polymerase chain reaction, expressed in Escherichia coli and purified by chromatography. Fab fragments of the parental antibody were prepared by papain digestion. Antibody fragments were formulated with a penetration and a viscosity enhancer and were applied to the surface of perfused pig corneas for up to 10 hours in vitro. Penetration was quantified by flow cytometry on rat thymocytes. RESULTS: 20-mer antibody fragments formed natural monomers and dimers following purification that could be separately isolated, while 11-mer fragments were dimeric. All formats of fragment (20-mer monomers and dimers, 11-mer dimers, Fab) showed penetration through the pig cornea after 6 hours of intermittent topical administration. CONCLUSION: Antibody fragments of different shapes and sizes can penetrate the cornea after topical administration, thereby increasing the potential of this class of proteins for topical ophthalmic use.en
dc.format.extent112181 bytes
dc.identifier.citationBrereton, H.M. Taylor, S D Farrall, A. Hocking, D. Thiel, M.A. Tea, M. Coster, D.J. Wililams, K.A. 2005. Influence of format on in vitro penetration of antibody fragments through porcine cornea. 'British Journal of Ophthalmology' Vol. 89, No. 9, 1205-1209en
dc.oaire.license.condition.licenseIn Copyright
dc.publisherBMJ Publishing Group -
dc.subjectAdministration, Topicalen
dc.subjectAntigens, CD4en
dc.subjectEpithelium, Cornealen
dc.subjectEscherichia colien
dc.subjectFlow Cytometryen
dc.subjectImmunoglobulin Fab Fragmentsen
dc.subjectImmunoglobulin Variable Regionen
dc.subjectResearch Support, Non-U.S. Gov'ten
dc.subjectTissue Culture Techniquesen
dc.subject.otherAustralian Standard Research Classification 321016 Ophthalmology and Vision Scienceen
dc.titleInfluence of format on in vitro penetration of antibody fragments through porcine corneaen
local.contributor.authorOrcidLookupTaylor, Susan D:
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