An improved sandwich ELISA method for the determination of immunoreactive schizophyllan (SPG).

As it is important to determine the optimal serum concentration of schizophyllan (SPG) when it is used as an anti-cancer drug, we devised a solid-phase ELISA. We also developed a sandwich ELISA using murine anti-SPG monoclonal antibody as the first antibody and rabbit anti-SPG serum as the second antibody in order to improve the detection sensitivity. This assay was able to determine SPG concentrations over 1.0 ng/ml and the absorbance at 490 nm was directly proportional to the SPG concentration. SPG in rabbit serum, obtained after intravenous and intramuscular injection (SPG; 10 mg/kg), was determined by this sandwich ELISA. Furthermore, the sensitivity of this ELISA method was compared with that of the Limulus test. The Limulus test is able to detect SPG in physiological saline (pH 6.3) at concentrations greater than 1.0 microgram/ml, but the sensitivity increased when SPG was dissolved in alkaline solution (pH 12.0), enabling SPG to be measured almost down to 1.0 ng/ml. These data suggest that our sandwich ELISA may be used for the measurement of SPG in blood or tissue.