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 Fe(2+)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.
| Original language | English |
|---|---|
| Pages (from-to) | 583-592 |
| Number of pages | 10 |
| Journal | mAbs |
| Volume | 11 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - 3 Apr 2019 |
Funding
We thank the Kimmelman Center for Biomolecular Structure and Assembly at the Weizmann Institute of Science for its generous support (to M.S.). M. S. holds the Katzir-Makineni chair in chemistry. D. D. thanks the Israel Science Foundation and the Russell Berrie Nanotechnology Institute, Technion for their support. G. P. thanks Prof. M. Firer for the use of the ELISA reader and Ariel University for their support.