Purification of antibody fragments via interaction with detergent micellar aggregates

Gunasekaran Dhandapani, Ellen Wachtel, Ishita Das, Mordechai Sheves, Guy Patchornik

Research output: Contribution to journalArticlepeer-review

3 Scopus citations


The research described in this report seeks to present proof-of-concept for a novel and robust platform for purification of antibody fragments and to define and optimize the controlling parameters. Purification of antigen-binding F(ab′)2 fragments is achieved in the absence of chromatographic media or specific ligands, rather by using clusters of non-ionic detergent (e.g. Tween-60, Brij-O20) micelles chelated via Fe2+ ions and the hydrophobic chelator, bathophenanthroline (batho). These aggregates, quantitatively capture the F(ab′)2 fragment in the absence or presence of E. coli lysate and allow extraction of only the F(ab′)2 domain at pH 3.8 without concomitant aggregate dissolution or coextraction of bacterial impurities. Process yields range from 70 to 87% by densitometry. Recovered F(ab′)2 fragments are monomeric (by dynamic light scattering), preserve their secondary structure (by circular dichroism) and are as pure as those obtained via Protein A chromatography (from a mixture of F(ab′)2 and Fc fragments). The effect of process parameters on Ab binding and Ab extraction (e.g. temperature, pH, ionic strength, incubation time, composition of extraction buffer) are reported, using a monoclonal antibody (mAb) and polyclonal human IgG’s as test samples.

Original languageEnglish
Article number11697
JournalScientific Reports
Issue number1
StatePublished - Dec 2021


Dive into the research topics of 'Purification of antibody fragments via interaction with detergent micellar aggregates'. Together they form a unique fingerprint.

Cite this