Abstract
A method suitable for the isolation of monoclonal antibodies (Mabs) is described. The protocol utilizes a zirconia based column modified with ethylenediamine-N,N'-tetra(methylenephosphonic) acid to create a novel cation-exchange chromatographic support. Initial experiments using a linear salt gradient demonstrate the ability of this support to efficiently separate Mab from transferrin and bovine serum albumin in a model matrix. Results of the purification of Mab from an actual cell culture supernatant over a range in protein concentrations are also shown. Analyses by enzyme-linked immunosorbent assay and gel electrophoresis demonstrate that Mabs can be recovered from a cell culture supernatant at high yield (92-98%) and high purity (>95%) in a single chromatographic step. Copyright (C) 1999 Elsevier Science B.V.
Original language | English (US) |
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Pages (from-to) | 63-72 |
Number of pages | 10 |
Journal | Journal of Chromatography A |
Volume | 831 |
Issue number | 1 |
DOIs | |
State | Published - Jan 22 1999 |
Bibliographical note
Funding Information:This work was performed through a grant from the National Institutes of Health (GM 45988-05).
Keywords
- Monoclonal antibodies
- Zirconia