Analysis of oral carcinoma antigens by monoclonal antibodies

Abstract

Monoclonal antibodies against human oral squamous cell carcinoma transplanted into nude mice were produced and a λ gt11 cDNA library was constructed from its mRNA extracts. Carcinoma homogenate was used as immunogen and electrofusion was carried out with pulses of 1.0-3.5 kV/cm. Antibody production was immunohistochemically screened by treating sections of transplanted tumor with a complex mixture consisting of three components; hybridoma supernatant, biotin-labeled anti-mouse Igs, and normal mouse serum. Extracted mRNA was used to prepare an oligo dT primed cDNA library. One hundred percent hybridoma formation was observed at certain pulse amplitudes and 2.3% of the hybridoma immunoreacted with the transplanted tumor. A total of 1.2×10^7 independent clones were obtained in the library and the insert sizes were 0.6-1.6 kbp. These techniques may provide pathologists with new methods of obtaining valuable diagnostic markers.