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Techniques
Assessment of Southern Bean Mosaic Virus Monoclonal Antibodies for Affinity Chromatography. W. P. Ronald, Technician, Agriculture Canada, Research Station, 6660 N.W. Marine Drive, Vancouver, B.C., Canada V6T 1X2; J. H. Tremaine(2), and D. J. MacKenzie(3). (2)(3)Research scientist, and technician, respectively, Agriculture Canada, Research Station, 6660 N.W. Marine Drive, Vancouver, B.C., Canada V6T 1X2. Phytopathology 76:491-494. Accepted for publication 19 December 1985. Copyright 1986 Department of Agriculture, Government of Canada. DOI: 10.1094/Phyto-76-491.
The dissociation of immunoprecipitates of three monoclonal antibodies (B5, B6, and B10) with southern bean mosaic virus was assessed at pH values between 7.0 and 2.25 by density gradient analysis and by indirect ELISA. Both methods showed that B10 did not dissociate appreciably, B5 dissociated at pH 2.75-2.25, and B6 dissociated at pH 3.5-2.25. Purified virus was recovered from a B6 affinity column by pH gradient elution at pH 3.4-2.2 or by elution of virus swollen with 2 mM EDTA in pH 7 buffered saline. Virus partially purified by polyethylene glycol or by chromatography on Sephacryl S-300 was recovered by pH gradient elution, but clarified sap of infected plants destroyed the reactivity of the B6 affinity column. Electron microscopy showed virus particles on B6 agarose beads at higher pH values and the release of virus into lower pH washes.
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