A general platform for antibody purification utilizing engineered-micelles |
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Authors: | Gunasekaran Dhandapani Assaf Howard Thien Van Truong Thekke V. Baiju Ellina Kesselman Noga Friedman |
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Affiliation: | 1. Department of Chemical Sciences, Ariel University, Ariel, Israel;2. Department of Chemistry, Indian Institute of Technology Bombay, Powai, India;3. Faculty of Biotechnology and Food Engineering, Technion, Haifa, Israel;4. Faculty of Chemistry, Weizmann Institute of Science, Rehovot, Israel |
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Abstract: | We introduce a new concept and potentially general platform for antibody (Ab) purification that does not rely on chromatography or specific ligands (e.g., Protein A); rather, it makes use of detergent aggregates capable of efficiently capturing Ab while rejecting hydrophilic impurities. Captured Ab are then extracted from the aggregates in pure form without co-extraction of hydrophobic impurities or aggregate dissolution. The aggregates studied consist of conjugated “Engineered-micelles” built from the nonionic detergent, Tween-20; bathophenanthroline, a hydrophobic metal chelator, and Fe2+ions. When tested in serum-free media with or without bovine serum albumin as additive, human or mouse IgGs were recovered with good overall yields (70–80%, by densitometry). Extraction of IgGs with 7 different buffers at pH 3.8 sheds light on possible interactions between captured Ab and their surrounding detergent matrix that lead to purity very similar to that obtained via Protein A or Protein G resins. Extracted Ab preserve their secondary structure, specificity and monomeric character as determined by circular dichroism, enzyme-linked immunosorbent assay and dynamic light scattering, respectively. |
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Keywords: | Antibody purification IgG Protein A Protein G Chromatography |
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