P53 fusion protein expression in prokaryote and preparation of monoclonal antibody to P53

Objective: Conventional immunohistochemistry (IHC) is available to assess P⁵³ mutations, and expensive imported anti-P⁵³ monoclonal antibody has been used in China, it is necessary to study a new monoclonal antibody. Methods: The P⁵³ DNA fragment enconding N-terminal 180 amiao acide was obtained by PCR and was cloned into P^GEX-2T plasmid expressing glutathione S-transferase (GST). The P⁵³-GST fusion protein expressed by JM109 was used for immunizing BALB/C mice. We have raised one hybridoma strain secreting McAb to human P⁵³ (named M126). Results: The IHC analysis of 52 paraffin-embedded sections from human breast cancer with M126 and P_AB1801 (Zymed Co.) has showed that the positive immunoreactions were 25 cases (48%) and 22 cases (42.3%) respectively. The staining of M126 was stronger and preferable to P_AB1801. Conclusion: M126 can be instead of P_AB1801 for studying immunohistochemical analysis on P⁵³ protein.