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) |
|---|---|
| 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 |
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Keywords
- Monoclonal antibodies
- Zirconia
ASJC Scopus subject areas
- Analytical Chemistry
- Biochemistry
- Organic Chemistry
Cite this
Purification of monoclonal antibodies from cell culture supernatants using a modified zirconia based cation-exchange support. / Clausen, Andrew M.; Subramanian, Anuradha; Carr, Peter W.
In: Journal of Chromatography A, Vol. 831, No. 1, 22.01.1999, p. 63-72.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Purification of monoclonal antibodies from cell culture supernatants using a modified zirconia based cation-exchange support
AU - Clausen, Andrew M.
AU - Subramanian, Anuradha
AU - Carr, Peter W.
PY - 1999/1/22
Y1 - 1999/1/22
N2 - 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.
AB - 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.
KW - Monoclonal antibodies
KW - Zirconia
UR - http://www.scopus.com/inward/record.url?scp=0032923637&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0032923637&partnerID=8YFLogxK
U2 - 10.1016/S0021-9673(98)00999-6
DO - 10.1016/S0021-9673(98)00999-6
M3 - Article
C2 - 10028281
AN - SCOPUS:0032923637
VL - 831
SP - 63
EP - 72
JO - Journal of Chromatography A
JF - Journal of Chromatography A
SN - 0021-9673
IS - 1
ER -