Improved immunodetection of human papillomavirus E7

Human papillomavirus E7 (HPV E7) is a viral on-coprotein that plays an important role in cervical carcinogenesis through binding with retinoblas-toma protein (Rb). Inactivation of Rb by E7 is necessary but not sufficient for cellular trans-formation, suggesting other protein-protein inter-actions are required for E7-mediated celular trans-formation aside from the interaction with Rb. HPV E7 have been limited by its poor imuno-reactivity. In this report, we show that the fixation of purified recombinant HPV E7 on blotted nitro-celulose membrane with glutaldehyde markedly enhanced the imunoreactivity of HPV E7 protein. Using HeLa and Caski cel line which are infected with HPV 18 and HPV 16, respectively, we de-monstrated that native HPV E7 proteins also could be detected by this method. These results there-fore can provide the experimental conditions for detection of HPV E7 proteins with greater sensi- KCI Citation Count: 7