Single chain antibody fragments for ocular use produced at high levels in a commercial wheat variety

H.M. Brereton; D. Chamberlain; R. Yang; M. Tea; S. McNeil; D.J. Coster; K.A. Williams

doi:10.1016/j.jbiotec.2007.01.008

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Single chain antibody fragments for ocular use produced at high levels in a commercial wheat variety

Journal article

Peer reviewed

Single chain antibody fragments for ocular use produced at high levels in a commercial wheat variety

H.M. Brereton, D. Chamberlain, R. Yang, M. Tea, S. McNeil, D.J. Coster and K.A. Williams

Journal of Biotechnology, Vol.129(3), pp.539-546

01/05/2007

DOI: https://doi.org/10.1016/j.jbiotec.2007.01.008

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Abstract

We are investigating the use of single chain antibody fragments (scFv) in eye drops for diagnosis and treatment of eye diseases. For ocular use, recombinant proteins must be free of bacterial endotoxin that causes inflammation in the eye. We required a means of generating high yields of scFvs with little endotoxin contamination. Using microprojectile bombardment we produced transgenic lines of the commercial wheat variety, Westonia, that express two scFvs that bind to CD4 or CD28 on the surface of rat thymocytes. A high level of expression of active scFv in the range 50-180 μg/g was measured by quantitative flow cytometry in crude extracts made from mature seeds. The levels of expression were stable over four generations of transgenic plants and mature seeds were stored for one year with little loss of scFv activity. Substantial purification of scFv was achieved by immobilised metal affinity chromatography. Compared to bacterial extracts, crude transgenic seed extracts contained only a small amount of endotoxin (150 EU/ml) that will be easily removed by purification. The transgenic wheat lines express functional scFv at levels comparable to production in bacteria and promise to be superior to bacteria for production of scFv pharmaceuticals for ocular use.

Details

Title: Single chain antibody fragments for ocular use produced at high levels in a commercial wheat variety
Authors/Creators: H.M. Brereton (Author/Creator) - Flinders University
D. Chamberlain (Author/Creator) - Murdoch University
R. Yang (Author/Creator) - Murdoch University
M. Tea (Author/Creator) - Flinders University
S. McNeil (Author/Creator) - Murdoch University
D.J. Coster (Author/Creator) - Flinders University
K.A. Williams (Author/Creator) - Flinders University
Publication Details: Journal of Biotechnology, Vol.129(3), pp.539-546
Publisher: Elsevier BV
Identifiers: 991005541587207891
Copyright: © 2007 Elsevier B.V.
Murdoch Affiliation: State Agricultural Biotechnology Centre
Language: English
Resource Type: Journal article

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Collaboration types: Domestic collaboration
Citation topics: 3 Agriculture, Environment & Ecology; 3.267 Virology - Plant; 3.267.2266 Molecular Farming
Web Of Science research areas: Biotechnology & Applied Microbiology
ESI research areas: Biology & Biochemistry