Among the more than 120 different types of human papillomavirus (HPV), types 16 and 18 have been known to be high risk agents that cause cervical cancer. We examined, in an immuno-chromatographic analysis, the potential of using the early gene product. E7 protein, as a diagnostic marker of cervical cancer caused by HPV. We developed monoclonal antibodies specific to HPV-16 and 18 E7 proteins that were produced from bacterial cells using, gene recombinant technology. For each E7 protein, the optimal antibody pair was selected using the immuno-chromatographic sandwich-type binding system based on the lateral flow through membrane pores. Under these conditions, this rapid testing assay had a detection capability as low as 2 ng/mL of E7 protein. Furthermore, since viral analysis required the host cell to be lysed using chemicals such as detergents, it was possible that the E7 protein was structurally damaged during this process, which would result in a decrease in detection sensitivity. Therefore, we examined the detrimental effects caused by different detergents on the E7 protein using HeLa cells as the host. In these experiments, we found that the damage caused by the detergent, nonylphenylpolyethylene glycol (NP-40), was minimal relative to Triton X-100 commonly used or the cell lysis. Temperature also affected the stability oldie E7 protein, and we found that the E7 protein was stabilized at 4 T. for about 2 h, which was 4 times longer than at room temperature. Finally, a HPV-infected cervical cancer cell line, which was used as a real sample model, was treated using the optimized conditions and the presence of E7 proteins were analyzed by immuno-chromatography. The results of this experiment demonstrated that this rapid test could specifically detect HPV-infected samples.Cho IH, 2009, ANAL CHIM ACTA, V632, P247, DOI 10.1016/j.aca.2008.11.019Liang YJ, 2008, INT J BIOCHEM CELL B, V40, P2431, DOI 10.1016/j.biocel.2008.04.003Ressler S, 2007, CLIN CANCER RES, V13, P7067, DOI 10.1158/1078-0432.CCR-07-1222Jeon JH, 2007, EXP MOL MED, V39, P621Mirecka EA, 2006, PROTEIN EXPRES PURIF, V48, P281, DOI 10.1016/j.pep.2006.04.017Nishimura A, 2006, MICROBES INFECT, V8, P984, DOI 10.1016/j.micinf.2005.10.015Cho JH, 2006, ANAL CHEM, V78, P793, DOI 10.1021/ac051453vHuh KW, 2005, P NATL ACAD SCI USA, V102, P11492, DOI 10.1073/pnas.0505337102Ono T, 2003, J IMMUNOL METHODS, V272, P211DeFilippis RA, 2003, J VIROL, V77, P1551, DOI 10.1128/JVI.77.2.1551-1563.2003Jeon JH, 2002, EXP MOL MED, V34, P496VINCE A, 2002, J CLIN VIROL, V25, P109Chen XJS, 2001, J MOL BIOL, V307, P173Paek SH, 2000, METHODS, V22, P53, DOI 10.1006/meth.2000.1036Walboomers JMM, 1999, J PATHOL, V189, P12Wertlake P, 1999, J REPROD MED, V44, P11JEON S, 1995, P NATL ACAD SCI USA, V92, P1654WILBUR DC, 1994, AM J CLIN PATHOL, V101, P209SCHEFFNER M, 1993, CELL, V75, P495SELVEY LA, 1992, J VIROL METHODS, V37, P119MAY K, 1991, AM J OBSTET GYNECOL, V165, P2000DYSON N, 1989, SCIENCE, V243, P934NELSON PH, 1989, EMBO J, V8, P3905GISSMANN L, 1986, VIRAL ETIOLOGY CERVI, P217CHANG JY, 1985, EUR J BIOCHEM, V151, P217
