Characterization and Selection of 3-(1-Naphthoyl)-Indole Derivative-Specific Alpaca VHH Antibodies Using a Phage Display Library.

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.

Development and Characterization of Monoclonal Antibodies Specific for 3-(1-naphthoyl) Indole Derivatives.

3-(1-naphthoyl) indole is one of the raw materials that synthesizes a synthetic cannabinoid such as 1-pentyl-3-(1-naphthoyl) indole (JWH-018) and 1-butyl-3-(1-naphthoyl) indole (JWH-073). It is important to detect the 3-(1-naphthoyl) indole derivatives rapidly, sensitively, and comprehensively. We developed two monoclonal antibodies (MAb) against 3-(1-naphthoyl) indole derivatives, named NT1 (IgG1) and NT2 (IgG1), which were possibly effective for detecting 3-(1-naphthoyl) indole derivatives. The cross-reactive ability of these MAbs was evaluated using a competitive enzyme-linked immunosorbent assay (ELISA). In the results, we found both of these antibodies recognize 3-(1-naphthoyl) indole and its derivatives. However neither of these antibodies recognize naphtoic acid, 4-methyl-naphtoic acid, and indole. Sixty to 100 nanomole per liter of 3-(1-naphthoyl) indole derivatives, such as 1-methyl-3-(1-naphthoyl) indole, 1-ethyl-3-(1-naphthoyl) indole, and 1-octyl-3-(1-naphthoyl) indole, can be detected using both of the obtained MAbs. Thus, the MAbs produced in this study could be a useful tool for the detection of 3-(1-naphthoyl) indole derivatives.

New monoclonal antibodies specific for 1-(5-fluoropentyl)-3-(2-iodobenzoyl)indole.

1-(5-fluoropentyl)-3-(2-iodobenzoyl)indole (AM694) is one of the synthetic cannabinoids and an illegal drug in Japan. It is important to generate a monoclonal antibody (MAb) against AM694 for use in the rapid and sensitive detection of the drug. Two monoclonal antibodies, named HN0124 (IgG1) and NK0504 (IgG1), were obtained, which were possibly effective for detecting AM694 and its derivatives. The cross-reactive ability of these MAbs was evaluated using a competitive enzyme-linked immunosorbent assay. In the results, both of these antibodies recognize 1-(5-fluoropentyl)-3-(2-iodobenzoyl)indole, 1-(5-fluoropentyl)-3-(3-iodobenzoyl)indole, 1-(5-fluoropentyl)-3-(4-iodobenzoyl)indole. Forty nmol/L AM694 can be detected using HN0124 MAb. Thus, MAbs produced in this study could be considered a useful tool for the detection of AM694.

A novel monoclonal antibody specific for cocaine.

Detection systems for the illegal drug cocaine need to have a high sensitivity and specificity for cocaine and to be relatively easy to use. In the current study, a monoclonal antibody (MAb) with a high specificity for cocaine was produced. Enzyme-linked immunosorbent assay and fluorescence quenching immunoassay were used to screen the hybridomas. The MAb S27Y (IgG1) was shown to be sensitive and specific for cocaine and quenched fluorescence. Thus, S27Y has the potential to be used in screening assays for the rapid and sensitive detection of cocaine.