title: Fibulin-1 purification from human plasma using affinity chromatography on Factor H-Sepharose.
abstract: A method is reported to purify Fibulin-1 from human plasma resulting in a 36% recovery. The steps involve removal of the cryoglobulin and the vitamin K dependent proteins followed by polyethylene glycol and ammonium sulfate precipitations, DEAE-Sephadex column chromatography and finally Factor H-Sepharose affinity purification. The procedure is designed to be integrated into an overall scheme for the isolation of over 30 plasma proteins from a single batch of human plasma. Results from mass spectroscopy, SDS-PAGE, and Western blotting indicate that human plasma Fibulin-1 is a single chain of the largest isotype. Functional binding assays demonstrated calcium ion dependent interaction of Fibulin-1 for fibrinogen, fibronectin, and Factor H. The procedure described is the first to our knowledge that enables a large scale purification of Fibulin-1 from human plasma.
authors.affiliation: Torrey Pines Institute for Molecular Studies, 3550 General Atomics Court, San Diego, CA 92121, United States. Electronic address: rdiscipio@sbpdiscovery.org.
authors.forename: Richard G
authors.initials: RG
authors.lastname: DiScipio
authors.affiliation: Sanford-Burnham Prebys Medical Discovery Institute, 10901 North Torrey Pines RD La Jolla, CA 92037, United States.
authors.forename: Robert C
authors.initials: RC
authors.lastname: Liddington
authors.affiliation: Torrey Pines Institute for Molecular Studies, 3550 General Atomics Court, San Diego, CA 92121, United States.
authors.forename: Ingrid U
authors.initials: IU
authors.lastname: Schraufstatter
mesh_terms: D000645:Ammonium Sulfate / Q000737:chemistry; D002135:Calcium-Binding Proteins / Q000097:blood* / Q000737:chemistry; D002846:Chromatography, Affinity; D017242:Complement Factor H / Q000737:chemistry*; D005340:Fibrinogen / Q000737:chemistry; D005353:Fibronectins / Q000737:chemistry; D006801:Humans; D013058:Mass Spectrometry; D012685:Sepharose / Q000737:chemistry
chemical_list: D002135:Calcium-Binding Proteins; D005353:Fibronectins; C060820:fibulin; D017242:Complement Factor H; D005340:Fibrinogen; D012685:Sepharose; D000645:Ammonium Sulfate
keywords: Affinity chromatography; Complement Factor H; Fibulin-1; Human plasma; Purification
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