title: Identification and analysis of anti-HDL scFv-antibodies obtained from phage display based synthetic antibody library.
abstract: OBJECTIVE
  In epidemiological studies plasma high density lipoprotein cholesterol (HDL-C) levels are found to correlate inversely with atherosclerotic cardiovascular events. HDL consists of different subpopulations and they vary in their anti-atherogenic properties. The aim of this study is to isolate coronary artery disease (CAD) specific anti-HDL scFv-antibodies.
  
  
  METHODS
  To obtain CAD specific HDL binders, we used phage displayed synthetic antibody libraries to enrich specific antibodies against HDL isolated from CAD patients. The antibodies were affinity purified. Their capability to recognize apolipoproteins A-I and A-II, various HDL forms differing in lipid/protein ratios and plasma HDL, was studied using time-resolved fluorescence based immunoassay.
  
  
  RESULTS
  Using different selection strategies and immunoassay based screening we obtained altogether 1200 clones displaying HDL binding activity. By sequencing 337, we identified 264 unique antibodies against HDL. A set of 61 antibodies were selected for further analysis. We found a variety of antibodies with different binding profiles, including apoA-I binding antibodies either in lipid-dependent or lipid-independent manner and binders against apoA-II. Several antibodies were able to discriminate between HDL derived from CAD patients and healthy controls. A majority of the antibodies were immunoreactive with HDL in plasma.
  
  
  CONCLUSIONS
  The novel HDL recognizing antibodies isolated from synthetic antibody phage library have displayed interesting HDL-binding characteristics suggesting that, in addition to use as research tools, a part of them might be useful for the development of diagnostic methods for CAD risk assessment.
authors.affiliation: Department of Biochemistry, Division of Biotechnology, University of Turku, Turku, Finland. Electronic address: priyanka.micro@gmail.com.
authors.forename: Priyanka
authors.initials: P
authors.lastname: Negi
authors.affiliation: Department of Biochemistry, Division of Biotechnology, University of Turku, Turku, Finland.
authors.forename: Janita
authors.initials: J
authors.lastname: Lövgren
authors.affiliation: Department of Biochemistry, Division of Biotechnology, University of Turku, Turku, Finland.
authors.forename: Päivi
authors.initials: P
authors.lastname: Malmi
authors.affiliation: Department of Biochemistry, Division of Biotechnology, University of Turku, Turku, Finland.
authors.forename: Nina
authors.initials: N
authors.lastname: Sirkka
authors.affiliation: Genomics and Biomarkers Unit, National Institute for Health and Welfare, Biomedicum, Helsinki, Finland.
authors.forename: Jari
authors.initials: J
authors.lastname: Metso
authors.affiliation: Department of Biochemistry, Division of Biotechnology, University of Turku, Turku, Finland.
authors.forename: Tuomas
authors.initials: T
authors.lastname: Huovinen
authors.affiliation: Department of Biochemistry, Division of Biotechnology, University of Turku, Turku, Finland.
authors.forename: Eeva-Christine
authors.initials: EC
authors.lastname: Brockmann
authors.affiliation: Department of Biochemistry, Division of Biotechnology, University of Turku, Turku, Finland.
authors.forename: Kim
authors.initials: K
authors.lastname: Pettersson
authors.affiliation: Genomics and Biomarkers Unit, National Institute for Health and Welfare, Biomedicum, Helsinki, Finland.
authors.forename: Matti
authors.initials: M
authors.lastname: Jauhiainen
authors.affiliation: Department of Biochemistry, Division of Biotechnology, University of Turku, Turku, Finland. Electronic address: urplammi@utu.fi.
authors.forename: Urpo
authors.initials: U
authors.lastname: Lamminmäki
mesh_terms: D001435:Bacteriophages / Q000235:genetics*; D016022:Case-Control Studies; D006801:Humans; D008075:Lipoproteins, HDL / Q000097:blood / Q000276:immunology*; D008297:Male; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology*
chemical_list: D008075:Lipoproteins, HDL; D057127:Single-Chain Antibodies
keywords: ApoA-I; ApoA-II; Apolipoproteins; CAD; Diagnostic tools; HDL; Lipoproteins