A non-chromatographic, ligand-free approach for antibody (Ab) purification, having the potential to replace the gold standard technology, i.e., Protein A chromatography, will be presented. Specially designed micellar aggregates, comprising of non-ionic detergents (e.g. Tween, Brij or Triton X-100), a hydrophobic chelator (e.g. bathophenanthroline) and metal cations (e.g. Fe2+), were found to lead to high Ab recovery yields (85-90%) and purity (>95%) while preserving Ab specificity and monomeric state. Interestingly, process efficiency is tightly correlated with micellar aggregate size (>1000 nm) as determined by dynamic light scattering (DLS) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. Moreover, purification is performed via three sequential filtration steps, implying that, the approach can be integrated into continuous production flows, where therapeutic grade monoclonal antibodies (mAbs) are purified at industrial-scales. Finally, the relevance of the same purification platform for IgA and IgM antibodies, will be discussed as well.

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Abstract Reference & Short Personal Biography of Presenting Author

Gunasekaran Dhandapani, Ariel University

Ph.D.: 2018-current in Chemistry and Biotechnology
Department of Chemical Sciences, Ariel University, Israel
MSc.: 2012-2014 in Genomics
School of Biological Sciences, Madurai Kamaraj University, India

B.Sc.: 2008-2012 in Bioinformatics
Department of Plant Molecular Biology & Bioinformatics Tamilnadu Agricultural University

Publications:

1. Dhandapani et al., Role of Amphiphilic [Metal:Chelator] Complexes in a Non-Chromatographic Antibody Purification Platform (Submitted).

2. Dhandapani et al., A general platform for antibody purification utilizing engineered-micelles, mAbs, (2019), 11:3, 583-592.

3. Dhandapani et al., Comparative proteome analysis reveals pathogen specific outer membrane proteins of Leptospira, Proteins: Structure, Function, and Bioinformatics (2018) 86, 712-722.

4. Dhandapani et al., Proteomic approach and expression analysis revealed the differential expression of predicted leptospiral proteases capable of ECM degradation, Biochimica et Biophysica Acta - Proteins and Proteomics (2018) 1866, 712-721.

5. Dhandapani et al., Molecular modeling and structural analysis of nAChR variants uncovers the mechanism of resistance to snake toxins, Journal of Biomolecular Structure and Dynamics, (2017), 35, 1654–1671.

6. Prachi Mehrotra, Gayatri Ramakrishnan, Gunasekaran Dhandapani, Narayanaswamy Srinivasan and Madathiparambil G. Madanan, Comparison of Leptospira interrogans and Leptospira biflexa genomes: analysis of potential leptospiral–host interactions, Molecular BioSystems (2017) 13, 883.