Purification of infectious canine parvovirus from cell culture by affinity chromatography with monoclonal antibodies |
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| Authors: | G F Rimmelzwaan J Groen N Juntti J S Teppema F G UytdeHaag A D Osterhaus |
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| Affiliation: | 1. Laboratory for Experimental Trauma Surgery, Justus-Liebig University Giessen, Kerkraderstr. 9, 35394 Giessen, Germany;2. Department of Diagnostic and Interventional Radiology, University Hospital of Giessen-Marburg, Klinikstrasse 33, 35392 Giessen, Germany;3. Laboratory for Experimental Radiology, Justus-Liebig University Giessen, Schubertstr. 81, 35392 Giessen, Germany;4. Institute of Orthopaedic Research and Biomechanics, Center of Musculoskeletal Research University of Ulm, Ulm, Germany;5. Department of Trauma Surgery Giessen, University Hospital of Giessen-Marburg, Campus: Giessen, Rudolf-Buchheim-Str. 7, 35392 Giessen, Germany;1. Center for Education and Research in Separation Methods & Bioanalytics BioSep, Faculty of Chemistry, Nicolaus Copernicus University, 7 Gagarin St., PL, 87 100 Toruń, Poland;2. Institute for Engineering of Polymer Materials and Dyes, 55 M. Sklodowskiej-Curie St., 87-100 Torun, Poland |
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| Abstract: | Immuno affinity chromatography with virus neutralizing monoclonal antibodies, directed to the haemagglutinating protein of canine parvovirus (CPV) was used to purify and concentrate CPV from infected cell culture. The procedure was monitored by testing the respective fractions in an infectivity titration system, in an ELISA, in a haemagglutination assay and by negative contrast electron microscopy to quantify CPV or CPV antigen. The degree of purification was further estimated by testing the fractions for total protein content in a colorimetric method, for bovine serum albumin content in an ELISA and by SDS-PAGE. Over 99% of the contaminating proteins proved to be removed, and 20% or 70-90% of infectious CPV or CPV antigen, respectively, was recovered. |
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