1. Characterization and Selection of 3-(1-Naphthoyl)-Indole Derivative-Specific Alpaca VHH Antibodies Using a Phage Display Library.
- Author
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Nakayama H, Murakami A, Yoshida M, Muraoka J, Wakai J, Kenjyou N, and Ito Y
- Subjects
- Animals, Antibody Specificity immunology, Antigens immunology, Enzyme-Linked Immunosorbent Assay, Indoles immunology, Peptide Library, Single-Domain Antibodies biosynthesis, Antibodies immunology, Camelids, New World immunology, Single-Domain Antibodies immunology
- Abstract
A new alpaca VHH antibody library against 3-(1-naphthoyl)-indole derivatives was developed from alpaca immunized with 7-(3-(1-naphthoyl)-1H-indol-1-yl)-heptanoic acid-keyhole limpet hemocyanin (Hep-KLH) protein conjugates as the immunogen. From this library, two 3-(1-naphthoyl)-indole derivative-specific clones, named NN01 and NN02, were isolated using biopanning technology. The binding specificity of these clones was confirmed using a competitive enzyme-linked immunosorbent assay (c-ELISA). Based on the results of c-ELISA, a median inhibitory concentration (IC50) of these two VHH antibodies, NN01 and NN02, in the case of 7-(3-(1-naphthoyl)-1H-indol-1-yl)-heptanoic acid (Hep; one of 3-(1-naphthoyl)-indole derivatives) as an inhibitor exhibited an approximate 3 × 10(-7) M and 6 × 10(-7) M, respectively. Thus, VHH antibodies produced in this study could be considered a useful tool for the detection of 3-(1-naphthoyl)-indole derivatives.
- Published
- 2016
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