Isolation and characterization of pharmaceutical grade human pentraxins, serum amyloid P component and C-reactive protein, for clinical use.

Abstract

The human pentraxin proteins, serum amyloid P component (SAP) and C-reactive protein (CRP) are important in routine clinical diagnosis, SAP for systemic amyloidosis and CRP for monitoring the non-specific acute phase response. They are also targets for novel therapies currently in development but their roles in health and disease are controversial. Thus, both for clinical use and to rigorously elucidate their functions, structurally and functionally intact, pharmaceutical grade preparations of the natural, authentic proteins are required. We report here the production from normal human donor plasma and the characterization of the first such preparations. Importantly, we demonstrate that, contrary to reports using recombinant proteins and less well characterized preparations, neither CRP nor SAP stimulate the release by human peripheral blood mononuclear cells in vitro of any TNFα, IL-6 or IL-8, nor does SAP cause release of IL-1β or IL-10. Furthermore neither of our preparations was pro-inflammatory in mice in vivo.

• Authors: Mark B Pepys, J Ruth Gallimore, Joanne Lloyd, Zhanhong Li, David Graham, Graham W Taylor, Stephan Ellmerich, Palma P Mangione, Glenys A Tennent, Winston L Hutchinson, David J Millar, Gary Bennett, John More, David Evans, Yogesh Mistry, Stephen Poole, Philip N Hawkins
• Journal: Journal of immunological methods (J Immunol Methods) Vol. 384 Issue 1-2 Pg. 92-102 (Oct 31 2012) ISSN: 1872-7905 [Electronic] Netherlands
• PMID: 22867744 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2012 Elsevier B.V. All rights reserved.

Chemical References

• Acute-Phase Proteins
• Cytokines
• Interleukin-1beta
• Interleukin-6
• Interleukin-8
• Serum Amyloid P-Component
• Tumor Necrosis Factor-alpha
• Interleukin-10
• C-Reactive Protein

Topics

• Acute-Phase Proteins (analysis, isolation & purification, pharmacology)
• Amyloidosis (blood)
• Animals
• C-Reactive Protein (analysis, isolation & purification, pharmacology)
• Cells, Cultured
• Chromatography, Gel
• Cytokines (metabolism)
• Electrophoresis, Polyacrylamide Gel
• Enzyme-Linked Immunosorbent Assay
• Female
• Humans
• Interleukin-10 (metabolism)
• Interleukin-1beta (metabolism)
• Interleukin-6 (metabolism)
• Interleukin-8 (metabolism)
• Leukocytes, Mononuclear (drug effects, metabolism)
• Mice
• Mice, Inbred C57BL
• Serum Amyloid P-Component (analysis, isolation & purification, pharmacology)
• Spectrometry, Mass, Electrospray Ionization
• Tumor Necrosis Factor-alpha (metabolism)