[Production and identification of the monoclonal antibodies specific for MHC class I complexes bound with HPV16E7 CTL...

To produce specific monoclonal-antibodies (MAbs) for MHC class I complexes bound with HPV16E7 CTL epitopic peptide (49-57), and provide a foundation for the investigation of the present pathway of viral antigen protein after in vitro viral infection.

Highly purified HPV16E7CTL epitope (49-57) (RAHYNIVTF) was produced, and then TAP deficient RMA-S cells incubated with RAHYNIVTF were used to immunize the BALB/c mouse. The spleen cells of the mice were regularly harvested and fused with the SP2/0 cells. The growing fusion wells were screened and the abstracted Mabs were identified in terms of sensitivity, specificity and affinity.

The screened hybriroma cells could steadily secrete the MAbs specific for MHC class I complexes bound with HPV16E7CTL epitopic peptide. The MAbs showed high reactivity with TAP-deficient RMA-S cells loaded with RAHYNIVTF and RMA-S cells which have the ability to process the endogenous MHC class I complexes, while minimally bound to class I molecules bearing other peptides, the results indicated excellent sensitivity, specificity and affinity of the MAbs.

The experiments provide a method for producing MAbs for epitopic peptide bound MHC class I complexes.