Purification Of Human Immunoglobulins A, G And M From Cohn ...

Full text links CiteDisplay options Display options Format AbstractPubMedPMID

Abstract

This work describes attempts to purify human IgG, IgA and IgM from Cohn fraction II/III using HWRGWV affinity peptide resin. The effects of peptide density and different elution additives on recovery of the three antibodies were investigated. At low peptide density, salting-in salts such as magnesium chloride and calcium chloride facilitated antibody elution. Ethylene glycol, urea and arginine also facilitated elution because of their ability to decrease hydrophobic interactions, hydrogen bonding and electrostatic interactions. However, at high peptide density, no recovery improvements were observed because of increased non-specific hydrophobic interactions. The final elution conditions for each antibody were chosen based on the resulting yields and purities when a 10:2:1mg/mL mixture of human IgG, IgA and IgM was used as starting material. Different pretreatment methods were employed in order to improve the purity of antibodies from Cohn fraction II/III. After pretreatment with caprylic acid precipitation or combination of caprylic acid and polyethylene glycol precipitation, purities over 95% and yields of about 60% were obtained for hIgG, which are comparable to current chromatographic purification methods involving two chromatography steps when hIgG is isolated from plasma fractions. A hIgA-enriched fraction with 42% hIgA and 56% hIgG, as well as a hIgM enriched fraction with 46% hIgM, 28% hIgA and 24% hIgG, were obtained as the by-products.

Copyright © 2012 Elsevier B.V. All rights reserved.

PubMed Disclaimer

MeSH terms

• Ammonium Sulfate / chemistry Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Arginine / chemistry Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Blood Proteins / chemistry* Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Caprylates / chemistry Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Chromatography, Affinity / methods* Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Ethylene Glycols / chemistry Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Humans Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Immunoglobulin Isotypes / blood Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Immunoglobulin Isotypes / isolation & purification* Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Peptides / chemistry Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Salts / chemistry Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search

Substances

• Blood Proteins Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Caprylates Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Cohn fraction II Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Cohn fraction III Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Ethylene Glycols Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Immunoglobulin Isotypes Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Peptides Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Salts Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Arginine Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• octanoic acid Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Ammonium Sulfate Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search

LinkOut - more resources

• Full Text Sources

  • Elsevier Science

• Other Literature Sources

  • The Lens - Patent Citations Database

• Miscellaneous

  • NCI CPTAC Assay Portal