MilliporeSigma

Conjugated detergent micelles as a platform for IgM purification.

Biotechnology and bioengineering (2022-03-25)
Gunasekaran Dhandapani, Ellen Wachtel, Ishita Das, Mordechai Sheves, Guy Patchornik
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
BRIJ® O20, average Mn ~1,150
Sigma-Aldrich
IgM from bovine serum, reagent grade, ≥95% (SDS-PAGE or HPLC), buffered aqueous solution
Sigma-Aldrich
IgM from human serum, reagent grade, ~95% (HPLC), buffered aqueous solution