title: 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.affiliation: Wolfson Drug Discovery Unit, Centre for Amyloidosis and Acute Phase Proteins, Division of Medicine, University College London, Rowland Hill Street, London NW3 2PF, UK. m.pepys@ucl.ac.uk
authors.forename: Mark B
authors.initials: MB
authors.lastname: Pepys
authors.forename: J Ruth
authors.initials: JR
authors.lastname: Gallimore
authors.forename: Joanne
authors.initials: J
authors.lastname: Lloyd
authors.forename: Zhanhong
authors.initials: Z
authors.lastname: Li
authors.forename: David
authors.initials: D
authors.lastname: Graham
authors.forename: Graham W
authors.initials: GW
authors.lastname: Taylor
authors.forename: Stephan
authors.initials: S
authors.lastname: Ellmerich
authors.forename: Palma P
authors.initials: PP
authors.lastname: Mangione
authors.forename: Glenys A
authors.initials: GA
authors.lastname: Tennent
authors.forename: Winston L
authors.initials: WL
authors.lastname: Hutchinson
authors.forename: David J
authors.initials: DJ
authors.lastname: Millar
authors.forename: Gary
authors.initials: G
authors.lastname: Bennett
authors.forename: John
authors.initials: J
authors.lastname: More
authors.forename: David
authors.initials: D
authors.lastname: Evans
authors.forename: Yogesh
authors.initials: Y
authors.lastname: Mistry
authors.forename: Stephen
authors.initials: S
authors.lastname: Poole
authors.forename: Philip N
authors.initials: PN
authors.lastname: Hawkins
mesh_terms: D000209:Acute-Phase Proteins / Q000032:analysis / Q000302:isolation & purification / Q000494:pharmacology; D000686:Amyloidosis / Q000097:blood; D000818:Animals; D002097:C-Reactive Protein / Q000032:analysis* / Q000302:isolation & purification / Q000494:pharmacology; D002478:Cells, Cultured; D002850:Chromatography, Gel; D016207:Cytokines / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D016753:Interleukin-10 / Q000378:metabolism; D053583:Interleukin-1beta / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D000683:Serum Amyloid P-Component / Q000032:analysis* / Q000302:isolation & purification / Q000494:pharmacology; D021241:Spectrometry, Mass, Electrospray Ionization; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D000209:Acute-Phase Proteins; D016207:Cytokines; D053583:Interleukin-1beta; D015850:Interleukin-6; D016209:Interleukin-8; D000683:Serum Amyloid P-Component; D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10; D002097:C-Reactive Protein