Engineered-Micelles: Alternative to Protein A Chromatography?

Gunasekaran Dhandapani, Chemical Sciences, Ariel University, Ariel, Israel (gunasekaran.vpm1990@gmail.com)

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Abstract Reference & Short Personal Biography of Presenting Author

We introduce a new concept and a potentially general platform for antibody (Ab) purification that does not rely on chromatography nor specific ligands (e.g. Protein A); rather, it makes use of detergent aggregates capable of efficiently capturing Ab's while rejecting hydrophilic impurities. Captured Ab's are then extracted from the aggregates in pure form (>95%) without co-extraction of hydrophobic impurities or aggregate dissolution. The aggregates studied consist of conjugated "Engineered-micelles" built from the nonionic detergent, Tween-20; the hydrophobic metal chelator, bathophenanthroline and Fe2+ions. When tested in serum-free media with\without BSA (or HSA) as additives, human or mouse IgG's were recovered at high overall yields (74-80%). Extraction of IgG's with 7 different buffers at pH 3.8 sheds light on possible interactions between captured Ab's and their surrounding detergent matrix. Extracted Ab's preserve their secondary structure, specificity and monomeric character as determined by CD, ELISA and DLS, respectively. Possible integration of the approach within industrial scale downstream processing of therapeutic grade monoclonal Ab's, will be  discussed.

Short prsonal biography: I graduated from the centre for excellence in Genomic Sciences, Madurai Kamaraj University, India. In 2017, I joined the Patchornik group at the Chemical Sciences deparment at Ariel Univeristy, Israel where we focuse on purification of soluble and membrane proteins via non-chromatographic strategies.