Table 1.

Correlation of patient's anti-p53 antibody production with p53 mutations and immunodominant epitopes

Patient no.Mutation codonp53 antibodyWestern blot fragments
40tyr→cys+2C, 23
53tyr→cys+NC, N5, 2C, 35, 23
168tyr→cys+N5, NC, 35
66arg→gln+5, NC, 23
226gly→glu+N5, NC
309pro→ser+N5, NC
296pro→leu+N5, 35
200his→arg-
192arg→gly-
69ile→asn-
245cys→tyr-
62G insert-
99arg→leu-
27tyr→ser-
71glu→stop-
  • NOTE: Patients' sera reacted with evolutionarily conserved domains of the p53 protein to determine possible immunodominant epitopes on the p53 molecule. Tumors from the patients were sequenced to determine the exact p53 mutation. Sera was tested for the presence of anti-p53 antibodies using a nondenaturing immunoprecipitation assay. Sera containing anti-p53 antibodies were incubated with various p53 fragments that were created using glutathione S-transferase fusion proteins in E. coli. This Western blot assay determined if the patient's sera bound to particular p53 fragments more frequently than others, thereby establishing if possible immunodominant portions7 of the p53 protein existed.