TY - JOUR
T1 - Conjugated detergent micelles as a platform for IgM purification
AU - Dhandapani, Gunasekaran
AU - Wachtel, Ellen
AU - Das, Ishita
AU - Sheves, Mordechai
AU - Patchornik, Guy
N1 - Publisher Copyright:
© 2022 The Authors. Biotechnology and Bioengineering published by Wiley Periodicals LLC.
PY - 2022/4/4
Y1 - 2022/4/4
N2 - Immunoglobulin M (IgM) antibodies hold promise as anticancer drugs and as agents for promoting immune homeostasis. This promise has not been realized due to low expression levels in mammalian cells producing IgM class antibodies, and the failure of protein A chromatography for IgM purification. Here, we describe a nonchromatographic platform for quantitatively capturing IgMs at neutral pH, which is then recovered with 86%–94% yield and >95% purity at pH 3. The platform contains micelles conjugated with the [(bathophenanthroline)3:Fe2+] amphiphilic complex. Inclusion of amino acid monomers, for example, phenylalanine or tyrosine, during conjugation of detergent micelles, allows subsequent extraction of IgMs at close to neutral pH. With the successful implementation of this purification platform for both polyclonal humans and bovine IgMs, we anticipate similar results for monoclonal IgMs, most relevant for the pharmaceutical industry.
AB - Immunoglobulin M (IgM) antibodies hold promise as anticancer drugs and as agents for promoting immune homeostasis. This promise has not been realized due to low expression levels in mammalian cells producing IgM class antibodies, and the failure of protein A chromatography for IgM purification. Here, we describe a nonchromatographic platform for quantitatively capturing IgMs at neutral pH, which is then recovered with 86%–94% yield and >95% purity at pH 3. The platform contains micelles conjugated with the [(bathophenanthroline)3:Fe2+] amphiphilic complex. Inclusion of amino acid monomers, for example, phenylalanine or tyrosine, during conjugation of detergent micelles, allows subsequent extraction of IgMs at close to neutral pH. With the successful implementation of this purification platform for both polyclonal humans and bovine IgMs, we anticipate similar results for monoclonal IgMs, most relevant for the pharmaceutical industry.
KW - IgM purification
KW - [metal:chelator] complexes
KW - conjugated micelles
KW - hydrophobic amino acids
KW - ligand free
KW - nonchromatographic
UR - http://www.scopus.com/inward/record.url?scp=85127396538&partnerID=8YFLogxK
U2 - 10.1002/bit.28089
DO - 10.1002/bit.28089
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C2 - 35324016
AN - SCOPUS:85127396538
SN - 0006-3592
VL - 119
SP - 1997
EP - 2003
JO - Biotechnology and Bioengineering
JF - Biotechnology and Bioengineering
IS - 7
ER -