PMID: 22570865May 10, 2012Paper

Recombinant IgA production: single step affinity purification using camelid ligands and product characterization

Journal of Immunological Methods
David ReinhartRenate Kunert

Abstract

Immunoglobulins of isotype A (IgA) mediate a key role in mucosal immunity and are promising new immunotherapeutic candidates, but difficulties in obtaining enough material often hamper their in vivo exploration. We established recombinant Chinese hamster ovary (CHO) cells which stably expressed two IgA1 antibodies under serum-free conditions. The two cell lines achieved significantly different specific productivities of 16 pg per cell and day and 100 times less, a common phenomenon in recombinant antibody expression which challenges the production and purification process. Polymeric IgA in crude culture supernatants was assembled with J chain and showed expected specificity. We employed an immobilized camelid anti-human alpha-chain VHH ligand and isolated both recombinant IgAs at high purity and yield in a single chromatographic step. The described method was irrespective of the light chain and specificity and may be used as a generic capture step for the isolation of any IgA. Results were compared with a multistep purification process consisting of an affinity step based on immobilized jacalin followed by anion exchange and hydrophobic interaction chromatography.

References

Jul 1, 1980·Proceedings of the National Academy of Sciences of the United States of America·G Urlaub, L A Chasin
Mar 17, 1999·Proceedings of the National Academy of Sciences of the United States of America·J BerdozB Corthésy
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May 25, 2010·Journal of Immunological Methods·Karola Vorauer-UhlRenate Kunert

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Citations

Jul 13, 2012·Journal of Translational Medicine·Nicolas RuffinRobin J Shattock
Feb 22, 2014·Applied Microbiology and Biotechnology·David ReinhartRenate Kunert
Dec 3, 2015·Biotechnology Advances·Nikolay VasilevAndreas Schiermeyer
Oct 30, 2015·Cellular and Molecular Life Sciences : CMLS·Vikram VirdiAnn Depicker
Jun 3, 2015·Journal of Separation Science·Gérald PerretEgisto Boschetti
Apr 26, 2015·Journal of Immunological Methods·Valérie Lorin, Hugo Mouquet
Mar 13, 2014·Cytotechnology·Veronika ChromikovaRenate Kunert
Sep 27, 2014·Biotechnology Letters·David Reinhart, Renate Kunert
Oct 21, 2016·Biotechnology Journal·Timothy M PabstAlan K Hunter
Jul 14, 2020·Frontiers in Bioengineering and Biotechnology·Steffen C HinzHarald Kolmar
Sep 28, 2017·Bioengineering·Felix HartSteffen Goletz

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