A comparative study of monoclonal antibodies (Mabs) purified from cell culture supernatant on EDTPA-modified zirconia beads and protein A-hyper D support

被引:1
|
作者
Subramanian, A
Martinez, B
Holm, J
Carr, PW
McNeff, CV
机构
[1] Univ Nebraska, Lincoln, NE 68588 USA
[2] Univ Minnesota, Dept Chem, St Paul, MN 55108 USA
[3] Zirchrom, Anoka, MN USA
关键词
zirconia; EPTPA; monoclonal antibody; biological activity; disassociation constant (K-d); protein-A;
D O I
10.1080/10826070500474055
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Colloidal zirconia was spray dried to yield zirconia particles, which were further modified with N,N,N',N'-Ethylenediamine tetra methylenephosphonic acid (EDTPA) to yield a support for use in bioseparations. EDTPA modified zirconia particles will be further referred to as, r_PEZ. Cell culture supernatants rich in monoclonal antibody (Mab) subtypes IgG(1), IgG(2a), IgG(2b), and IgG(3) were chromatographed on a r_PEZ column, and on a protein A-hyper D column that was purchased commercially. All Mab subtypes bound to r_PEZ and process yields in the range of 88 to 99% were obtained. The purity of the Mab products were ascertained by gel electrophoretic analysis and were estimated to be greater than 95%. The purified Mab products obtained from r_PEZ and protein A columns were compared to the reference Mab standard in biological and enzymatic assays. The value of the dissociation constant (K-d ) was found to be comparable and was in the range to that obtained with reference Mab standard (0.231 +/- 0.03 M). In addition, Mabs purified with r_PEZ had the same deglycosylation profile as the reference Mab standard. Thus, it appears that the r_PEZ purified Mab is similar in activity to Mab purified with a protein A support and in addition, the zirconia surface does not adversely impact the activity of the purified Mab.
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页码:471 / 484
页数:14
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