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Affinity chromatographic purification of human immunoglobulin M from human B lymphocyte cell culture supernatant

Paper ID Volume ID Publish Year Pages File Format Full-Text
3443 171 2013 8 PDF Available
Title
Affinity chromatographic purification of human immunoglobulin M from human B lymphocyte cell culture supernatant
Abstract

Compared to immunoglobulin G purification with extensively studied affinity ligands such as protein A and protein G, little work has been done on affinity chromatographic purification of immunoglobulin M. Hexamer peptide ligand HWRGWV, previously shown to bind specifically to the Fc fragment of IgG, also demonstrated potential for IgM purification. This study presents further characterization and investigation of this ligand for its potential for purification of IgM. Different running conditions were employed in order to improve the recovery and purity of IgM. The final recovery and purity of the antibody is feedstock dependent, but can reach levels of both recovery and purity as high as 95%. The dependence of the recovery and purity on total loading amount and initial IgM concentration were investigated and discussed. Although relatively low dynamic binding capacities (DBC) in the range of 4.6–13.1 mg IgM/mL resin at linear flow rates from 173 to 35 cm/h were obtained for IgM compared to IgG because of the large molecular weight of IgM, the DBC value of HWRGWV for IgM is much greater than protein-based IgM affinity ligands found in the literature and is competitive with current commercially available affinity ligands, such as KAPTIVE-M, CaptureSelect IgM and Ultralink Immobilized Mannan Binding Protein.

► HWRGWV demonstrates great potential for affinity purification of hIgM. ► DBC for hIgM with HWRGWV is highly dependent on linear flow rate. ► Final recovery and purity for hIgM purification is feedstock dependent. ► As high as 95% for both recovery and purity can be achieved for hIgM purification.

Keywords
Affinity chromatography; Hexamer peptide; Human immunoglobulin; IgM; Dynamic binding capacity; Elution additive
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Affinity chromatographic purification of human immunoglobulin M from human B lymphocyte cell culture supernatant
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 70, 15 January 2013, Pages 63–70
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us