Selection of antibody fragments specific for an alpha-helix region of acylphosphatase |
| |
Authors: | Degl'Innocenti Donatella Taddei Niccolò Ramazzotti Matteo Stefani Massimo Chiti Fabrizio Ramponi Giampietro |
| |
Institution: | Dipartimento di Scienze Biochimiche, Università degli Studi di Firenze, Viale Morgagni 50, Firenze 50134, Italy. donatclla.deglinnocenti@unifi.it |
| |
Abstract: | The native state of common-type acylphosphatase (AcP) elicits two alpha-helices spanning residues 22-32 and 55-67 in the protein sequence. A peptide corresponding to the second alpha-helix (helix-2) of the protein was used to select phage antibodies consisting of a single chain fragment variable. The selection was performed in the presence of trifluoroethanol, a cosolvent known to induce the formation of helical structure in peptides and proteins. Phage scFv antibodies capable of binding the peptide specifically in a trifluoroethanol-induced alpha-helical conformation were isolated by affinity selection (biopanning). Some of these scFvs were also able to bind the native protein but not the peptide in a non-helical unstructured state. This indicates that the structural determinant recognized by the selected antibodies is the alpha-helical conformation of this specific region, rather than simply its amino acid sequence. This study shows that phage display libraries can be used to raise antibodies one can use as reagents able to target regions of a protein with a specific native-like secondary structure. |
| |
Keywords: | scFv AcP peptide mimotope epitope binding helical structure biopanning |
本文献已被 PubMed 等数据库收录! |