Immunoreactivity of phage library-derived human single-chain antibodies to amyloid beta conformers in vitro
| Autor: | Takeshi Iwatsubo, Shinichiro Tsuyama, Tadafumi Hashimoto, Koichi Tanaka, Yuji Ito, Shuhei Hashiguchi, Sayaka Kuba, Kazuhisa Sugimura, Akifumi Kyuno, Toshihiro Nakashima, Sho Takiguchi, Tomoki Yoshihara |
|---|---|
| Rok vydání: | 2008 |
| Předmět: |
Amyloid beta-Peptides
Base Sequence Sequence Homology Amino Acid Amyloid beta Protein Conformation Molecular Sequence Data P3 peptide General Medicine Biopanning Biology Biochemistry Molecular biology Epitope Antibodies biology.protein Single-chain variable fragment Humans Bacteriophages Electrophoresis Polyacrylamide Gel Amino Acid Sequence Binding site Antibody Molecular Biology Single-Chain Antibodies DNA Primers |
| Zdroj: | Journal of biochemistry. 143(4) |
| ISSN: | 0021-924X |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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