Figure 1.

Strategy for the detection of tumor antigens from clinical biopsy samples. Cervical cancer biopsy material and blood were obtained from patients and processed in three ways: (1) Tumor DNA was extracted and the E7 oncogene sequenced to determine its conservation; (2) The protein fraction from the tumor was used to extract peptide–HLA complexes for MS³ analysis; and (3) RNA was isolated from patient PBMCs and used to obtain high resolution HLA typing via 454 sequencing. Collectively, the technologies allow DNA sequencing and T cell epitope predictions within conserved segments of tumor antigens bound to genotypically defined HLA molecules to be verified by MS³ analysis.