Biopolymers and cell. 2005; 21 (2): 180 - 183

Investigation of some structural-functional peculiarities of the immunogenic fragment Ki-67 antigen. Generation of polyclonal and monoclonal antibodies to Ki-67

I.O. Tykhonkova, V. V. Lisogubov, Yu. V. Khozhaenko, O. O. Dovgopola, S. I. Korovin, G. V. Ovcharenko, V. S. Usenko, M. V. Rodnin, V. V. Filonenko

The study of tumor-associated antigens function is a prerequisite for understanding the molecular mechanisms of oncogenesis. Earlier we have determined the nuclear protein Ki-67 as an autoantigen in human melanoma. cDNA of immunogenic fragment of Ki-67 has been cloned into pET23d vector and expressed in bacterial cells. The recombinant fragment of Ki-67 has been purified from the bacterial cell lysate has been used as an antigen for the production of polyclonal and monoclonal antibodies with following detection of Ki-67 in Breast adenocarcinomas. We have shown by the immunodot-blot analysis that the 18 kDa recombinant fragment of Ki-67 possesses a DNA-binding activity.

Key words: Ki-67 antigen, antibodies, DNA-binding activity, immunohistochemistry.