title: Search for amyloid-binding proteins by affinity chromatography.
abstract: 'Amyloid binging proteins' is a generic term used to designate proteins that interact with different forms of amyloidogenic peptides or proteins and that, as a result, may modulate their physiological and pathological functions by altering solubility, transport, clearance, degradation, and fibril formation. We describe a simple affinity chromatography protocol to isolate and characterize amyloid-binding proteins based on the use of sequential elution steps that may provide further information on the type of binding interaction. As an example, we depict the application of this protocol to the study of Alzheimer's amyloid β (Aβ) peptide-binding proteins derived from human plasma. Biochemical analysis of the proteins eluted under different conditions identified serum amyloid P component (SAP) and apolipoprotein J (clusterin) as the main plasma Aβ-binding proteins while various apolipoproteins (apoA-IV, apoE, and apoA-I), as well as albumin (HSA) and fibulin were identified as minor contributors.
authors.affiliation: Centro Nacional de Microbiología, Instituto de Salud Carlos III, Majadahonda, Madrid, Spain. mcalero@isciii.es
authors.forename: Miguel
authors.initials: M
authors.lastname: Calero
authors.forename: Agueda
authors.initials: A
authors.lastname: Rostagno
authors.forename: Jorge
authors.initials: J
authors.lastname: Ghiso
mesh_terms: D000595:Amino Acid Sequence; D016229:Amyloid beta-Peptides / Q000737:chemistry / Q000378:metabolism*; D001053:Apolipoproteins / Q000378:metabolism; D001798:Blood Proteins / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods*; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry / Q000378:metabolism; D008024:Ligands; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D015203:Reproducibility of Results; D012709:Serum Albumin / Q000378:metabolism
chemical_list: D016229:Amyloid beta-Peptides; D001053:Apolipoproteins; D001798:Blood Proteins; D002135:Calcium-Binding Proteins; D055767:Immobilized Proteins; D008024:Ligands; D010446:Peptide Fragments; D012709:Serum Albumin; C078592:amyloid beta-protein (1-40); C060820:fibulin