ENZYME-LINKED IMMUNOSORBENT ASSAYS (ELISA) FOR THE DETECTION OF ANTIBODIES TO UK114 AND UK150

Doses of UK101, a mammalian liver-extracted protein consisting of UK11 0, UK114 and UK150, were administered to advanced neoplastic patients to try a specific immunotherapy. Two ELISA immunoassays were developed in order to evaluate the antiUK114 and antiUK150 antibody titre. In b oth methods, we used the specific protein (UK114 or UK150) bound onto polystyrene microwells as catcher and a purified anti-human IgG (Fc) c onjugated with horseradish peroxidase as tracer. The enzymatic activit y bound onto solid phase was detected by TMB/H-2 O-2 chromogen/substra te system. A calibration curve was constructed by using a pool of UK10 1-treated patients' sera, which showed high concentrations of antiUK11 4 or antiUK150 antibodies. The antibody activity of each pool was expr essed in arbitrary unit (U/mL) and a panel of 86 normal human sera dra wn from hospital blood bank was used to define the cut-off values of t he assays. The evaluation of the antiUK114 antibody titre in 30 breast cancer patients and 31 colon cancer patients submitted to UK101 thera py is here reported. An increase of 63% and 87% was detected after 40 days of UK101 administration in the two groups of terminally ill patie nts, respectively.