Membrane chromatography for rapid purification of recombinant antithrombin III and monoclonal antibodies from cell culture supernatant

Lütkemeyer D, Bretschneider M, Büntemeyer H, Lehmann J (1993)
Journal of Chromatography, A 639(1): 57-66.

Download
OA
Journal Article | Published | English
Author
; ; ;
Abstract
The task of purifying monoclonal antibodies (MAbs) and human recombinant antithrombin Ill (rATIII) from cell culture supernatant was carried out using two different approaches, both based on the use of membraneous matrices. The first approach employed a strongly acidic and a strongly basic membrane ion exchanger, which were evaluated for their ability to purify monoclonal antibodies and the human active recombinant antithrombin III from cell culture supernatant. Within minutes gram amounts of product could be purified in a high-flux system, specially developed for this purpose, achieving purities of 80% for MAbs and 75% for rATIII, respectively. The capacity of the acidic membrane ion exchanger for MAbs was found to be 1 mg/cm2 with recoveries up to 96% and that of the basic membrane ion exchanger for rATIII was 0.15 mg/cm2 with recoveries up to 91%. The second approach consisted of using heparin, a mucopolysaccharide with a strong affinity towards ATIII, coupled to amine-modified or epoxy-activated membranes by reductive amination, for the purification of rATIII. The ATIII binding capacities of the membranes were found to be 91 mug/cm2 for the amine-modified and 39 mug/cm2 for the epoxy-activated membrane, achieving purities of 75%. The coupling proved to be fairly stable over a period of 5 months and the membranes remained operable even after steam sterilization and treatment with sodium dodecyl sulphate. Final purification in both instances was carried out by gel filtration.
Publishing Year
ISSN
PUB-ID

Cite this

Lütkemeyer D, Bretschneider M, Büntemeyer H, Lehmann J. Membrane chromatography for rapid purification of recombinant antithrombin III and monoclonal antibodies from cell culture supernatant. Journal of Chromatography, A. 1993;639(1):57-66.
Lütkemeyer, D., Bretschneider, M., Büntemeyer, H., & Lehmann, J. (1993). Membrane chromatography for rapid purification of recombinant antithrombin III and monoclonal antibodies from cell culture supernatant. Journal of Chromatography, A, 639(1), 57-66.
Lütkemeyer, D., Bretschneider, M., Büntemeyer, H., and Lehmann, J. (1993). Membrane chromatography for rapid purification of recombinant antithrombin III and monoclonal antibodies from cell culture supernatant. Journal of Chromatography, A 639, 57-66.
Lütkemeyer, D., et al., 1993. Membrane chromatography for rapid purification of recombinant antithrombin III and monoclonal antibodies from cell culture supernatant. Journal of Chromatography, A, 639(1), p 57-66.
D. Lütkemeyer, et al., “Membrane chromatography for rapid purification of recombinant antithrombin III and monoclonal antibodies from cell culture supernatant”, Journal of Chromatography, A, vol. 639, 1993, pp. 57-66.
Lütkemeyer, D., Bretschneider, M., Büntemeyer, H., Lehmann, J.: Membrane chromatography for rapid purification of recombinant antithrombin III and monoclonal antibodies from cell culture supernatant. Journal of Chromatography, A. 639, 57-66 (1993).
Lütkemeyer, Dirk, Bretschneider, M., Büntemeyer, Heino, and Lehmann, Jürgen. “Membrane chromatography for rapid purification of recombinant antithrombin III and monoclonal antibodies from cell culture supernatant”. Journal of Chromatography, A 639.1 (1993): 57-66.
Main File(s)
Access Level
OA Open Access

This data publication is cited in the following publications:
This publication cites the following data publications:

15 Citations in Europe PMC

Data provided by Europe PubMed Central.

A monolith purification process for virus-like particles from yeast homogenate.
Burden CS, Jin J, Podgornik A, Bracewell DG., J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 880(1), 2012
PMID: 22134039
Membrane-based techniques for the separation and purification of proteins: an overview.
Saxena A, Tripathi BP, Kumar M, Shahi VK., Adv Colloid Interface Sci 145(1-2), 2009
PMID: 18774120
Recovery of antithrombin III from milk by expanded bed chromatography.
Ozyurt S, Kirdar B, Ulgen KO., J Chromatogr A 944(1-2), 2002
PMID: 11831754
Affinity membrane chromatography for the analysis and purification of proteins.
Zou H, Luo Q, Zhou D., J. Biochem. Biophys. Methods 49(1-3), 2001
PMID: 11694281
Membrane ion-exchange chromatography for process-scale antibody purification.
Knudsen HL, Fahrner RL, Xu Y, Norling LA, Blank GS., J Chromatogr A 907(1-2), 2001
PMID: 11217020
Large-scale membrane adsorbers.
Demmer W, Nussbaumer D., J Chromatogr A 852(1), 1999
PMID: 10480231
Plasma protein fractionation with advanced membrane adsorbents.
Gebauer KH, Thommes J, Kula MR., Biotechnol. Bioeng. 54(2), 1997
PMID: 18634085

21 References

Data provided by Europe PubMed Central.

Surface-modified membranes as a matrix for protein purification.
Langlotz P, Kroner KH., J. Chromatogr. 591(1-2), 1992
PMID: 1613045

Kroner, DECHEMA Biotechnol. Conf. 5(), 1992

Champluvier, 1990

Briefs, DECHEMA Biotechnol. Conf. 4(), 1990

Kroner, Engineering Foundation Conference, Recovery of Biological Products VI, Interlaken, Switzerland (), 1992
Purification of enzymes by heparin-sepharose affinity chromatography.
Farooqui AA., J. Chromatogr. 184(3), 1980
PMID: 6997325
Binding of [3H]heparin to human plasma low density lipoprotein.
Pan YT, Kruski AW, Elbein AD., Arch. Biochem. Biophys. 189(2), 1978
PMID: 30401

Marshall, Biotechnology 11(7), 1989
Antithrombin III and heparin.
Barrowcliffe TW, Johnson EA, Thomas D., Br. Med. Bull. 34(2), 1978
PMID: 350337
The direct coupling of oligosaccharides to proteins and derivatized gels.
Gray GR., Arch. Biochem. Biophys. 163(1), 1974
PMID: 4859528
Improved method for the immobilization of heparin.
Sasaki H, Hayashi A, Kitagaki-Ogawa H, Matsumoto I, Seno N., J. Chromatogr. 400(), 1987
PMID: 3667742

Zettlmeissl, Biotechnology 5(), 1987
Optimization of serum-free fermentation processes for antibody production.
Buntemeyer H, Lutkemeyer D, Lehmann J., Cytotechnology 5(1), 1991
PMID: 1367052

Lütkemeyer, Bio Engineering 2(), 1992

Dubois, Anal. Chem. 28(), 1956

Bülles, 1990

Demmer, 1990

Export

0 Marked Publications

Open Data PUB

Web of Science

View record in Web of Science®

Sources

PMID: 8331144
PubMed | Europe PMC

Search this title in

Google Scholar