Immunoreactivity of phage library-derived human single-chain antibodies to amyloid beta conformers in vitro

J Biochem. 2008 Apr;143(4):475-86. doi: 10.1093/jb/mvm239. Epub 2008 Jan 2.

Abstract

The pathogenesis of Alzheimer's disease involves conformational changes of A beta. A series of antibodies recognizing a distinct conformation of A beta (snapshot antibody) is useful for both understanding the mechanism of molecular conversion and identifying diagnostic and therapeutic reagents. As A beta with various conformations can be prepared in vitro under varying physicochemical conditions, snapshot antibodies can be isolated by directly binding to target molecules with antibody-displaying phages. We tested the feasibility of this idea. We show a feature of several A beta-reactive antibodies isolated from our human single-chain Fv antibody-phage library and particularly report the characteristics of an scFv clone, B6, selected from the fibrillar A beta 1-42-coated biopanning. B6 bound to fibrillar A beta 1-42 as well as globulomer A beta 1-42 but not to soluble A beta 1-42 or A beta 1-40. B6 inhibited A beta 1-42 fibril formation with 600 nM IC50 in spite of being the monovalent scFv form. Epitope analysis suggested that the binding site might be located at the beta2 sheet of the C-terminus of A beta 1-42. Although it is believed that N-terminus-recognizing antibodies tend to show the capability to inhibit A beta 1-42 fibrillation, B6 is the first human inhibitory antibody recognizing the C-terminus of A beta 1-42.

MeSH terms

  • Amino Acid Sequence
  • Amyloid beta-Peptides / chemistry
  • Amyloid beta-Peptides / immunology*
  • Antibodies / genetics
  • Antibodies / immunology*
  • Bacteriophages / genetics*
  • Base Sequence
  • DNA Primers
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Molecular Sequence Data
  • Protein Conformation
  • Sequence Homology, Amino Acid

Substances

  • Amyloid beta-Peptides
  • Antibodies
  • DNA Primers