Takashi Usagawa

Preparation of monoclonal antibodies against okadaic acid prepared from the sponge Halichondria okadai

Three murine monoclonal antibodies, OA-1, OA-2 and OA-3, against okadaic acid were prepared from hybridoma clones obtained by fusion of mouse 653 myeloma cells with mouse immune spleen cells sensitized to okadaic acid-ovalbumin conjugate. Each antibody reacted with dinophysistoxin-1 ( = 35-methylokadaic acid) as well as okadaic acid, but did not react with the other diarrhetic shellfish poisons or related compounds, such as 7-O-palmitoyl-okadaic acid (analogue of dinophysistoxin-3), pectenotoxin-1 and yessotoxin. A competitive inhibition enzymelinked immunosorbent assay which employed OA-3 antibody was performed and showed a sensitivity of about 10 ppb (10 ng/ml) for okadaic acid. This simple and time-saving ELISA assay system may be useful for the specific detection of diarrhetic shellfish poisons.

Preparation of monoclonal antibodies against methamphetamine

15 monoclonal antibodies against methamphetamine (MA) were obtained from hybridoma clones by the fusion of mouse 653 myeloma cells with the spleen cells of the immunized mice. The monoclonal antibodies exhibited different cross-reactivities against MA-related compounds and one antibody (MA-15 Ab) was highly specific against MA. A competitive inhibition enzyme-linked immunosorbent assay was established and the detection limit for MA was approximately 20 ng/ml.

Characterization of morphine-specific monoclonal antibodies showing minimal cross-reactivity with codeine

Six murine monoclonal antibodies against morphine were produced using N-(4-aminobutyl)normorphine as a hapten. Most of the antibodies obtained distinguished the substituents at the 3 and 6 positions of morphine. This property of the antibodies led to a reduction in cross-reactivity with codeine, morphine-3-glucuronide (M-3-G) and morphine-6-glucuronide (M-6-G) to negligible levels. However, one of the antibodies distinguished the substituent only at the 3 position of morphine, which cross-reacted with M-6-G, naloxone and naltrexone. In the competitive inhibition enzyme-linked immunosorbent assay, morphine was detected at concentrations as low as circa 100 pg/ml.