Hessing, M., van Schijndel, H. B., van Grunsven, W. M., Wolf, Hans J. and Middeldorp, J. M.
(1992)
Purification and quantification of recombinant Epstein-Barr viral glycoproteins gp350/220 from Chinese hamster ovary cells.
Journal of chromatography 599 (1-2), pp. 267-272.
Date of publication of this fulltext: 11 Apr 2011 10:53at PubMed
Abstract
Truncated Epstein-Barr virus (EBV) membrane antigen gp350/220 (EBV-MA) lacking the membrane anchor was expressed and secreted into the medium of recombinant Chinese hamster ovary cells that had been cultured in Plasmapur hollow-fibre modules using defined serum-free medium. The EBV-MA in the medium was concentrated by 70% (w/v) ammonium sulphate precipitation and subsequently purified by ...
Abstract
Truncated Epstein-Barr virus (EBV) membrane antigen gp350/220 (EBV-MA) lacking the membrane anchor was expressed and secreted into the medium of recombinant Chinese hamster ovary cells that had been cultured in Plasmapur hollow-fibre modules using defined serum-free medium. The EBV-MA in the medium was concentrated by 70% (w/v) ammonium sulphate precipitation and subsequently purified by immunoaffinity chromatography using an anti-EBV-MA (EBV.0T6) monoclonal antibody (mAb) column. Adsorbed antigen was eluted with 3 M MgCl2 in phosphate-buffered saline, concentrated by Mono Q anion-exchange chromatography and analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, silver staining and Western blotting using EBV-positive serum and anti-EBV-MA specific mAbs. Monospecific polyclonal rabbit antibodies against the purified EBV-MA were raised and purified by protein G affinity chromatography. For the measurement of EBV-MA antigen levels a sandwich enzyme-linked immunosorbent assay using rabbit polyclonal antibodies and a horseradish peroxidase-conjugated anti-MA mAb was developed having a detection level of 10 ng/ml.
Export bibliographical data
| Item type: | Article |
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| Date: | 1992 |
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| Institutions: | Medicine > Lehrstuhl für Medizinische Mikrobiologie und Hygiene |
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| Identification Number: | | Value | Type |
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| 1320046 | PubMed ID |
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| Classification: | | Notation | Type |
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| Animals | MESH | | Antigens, Viral/isolation & purification | MESH | | Blotting, Western | MESH | | CHO Cells | MESH | | Cricetinae | MESH | | Electrophoresis, Polyacrylamide Gel | MESH | | Enzyme-Linked Immunosorbent Assay | MESH | | Herpesvirus 4, Human/immunology | MESH | | Recombinant Proteins/isolation & purification | MESH | | Viral Envelope Proteins/isolation & purification | MESH | | Viral Matrix Proteins | MESH |
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| Dewey Decimal Classification: | 600 Technology > 610 Medical sciences Medicine |
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| Status: | Published |
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| Refereed: | Unknown |
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| Created at the University of Regensburg: | Unknown |
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| Item ID: | 20424 |
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