Role of amphiphilic [metal:chelator] complexes in a non-chromatographic antibody purification platform.

Autor: Dhandapani G; Department of Chemical Sciences, Ariel University, 70400 Ariel, Israel., Nair DK; Department of Chemistry, Indian Institute of Technology Bombay, Powai, Mumbai 400076, India., Kale RR; Department of Chemistry, Indian Institute of Technology Bombay, Powai, Mumbai 400076, India., Wachtel E; Faculty of Chemistry, Weizmann Institute of Science, 76100 Rehovot, Israel., Namboothiri INN; Department of Chemistry, Indian Institute of Technology Bombay, Powai, Mumbai 400076, India., Patchornik G; Department of Chemical Sciences, Ariel University, 70400 Ariel, Israel. Electronic address: guyp@ariel.ac.il.
Jazyk: angličtina
Zdroj: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences [J Chromatogr B Analyt Technol Biomed Life Sci] 2019 Dec 01; Vol. 1133, pp. 121830. Date of Electronic Publication: 2019 Oct 21.
DOI: 10.1016/j.jchromb.2019.121830
Abstrakt: We have recently introduced a non-chromatographic alternative for antibody (Ab) purification, one which does not require the use of Protein A. With this approach, polyclonal human or mouse immunoglobulins (IgG's) are captured almost quantitatively by Tween-20 micelles conjugated with a [chelator:divalent metal cation] complex. Target IgG structure remains native even following extraction from the surfactant aggregate. In the present work, we explore the effect of varying both components of the [metal:chelator] pair on the yield of purified Ab. Capture efficiency is observed to correlate with the formation of sufficiently large detergent aggregates, as determined by dynamic light scattering (DLS) and polyacrylamide gel electrophoresis (PAGE). This, in turn, depends on the rigidity and aromaticity of the chelator. Detergent aggregates are stable over a wide range of pH values (pH = 3-9). Under acidic conditions (3-3.8) they lead to good IgG recovery yields (70-78%) with purity similar to that obtained with Protein A chromatography while maintaining the monomeric state of the IgG's. Finally, the influence of the environment and the presence of various water-soluble chelators (e.g. EDTA, histidine, imidazole) on process efficiency, is described.
(Copyright © 2019 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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