Generation of affibody molecules specific for HPV16 E7 recognition
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Xiangyang Xue1,*, Bingbing Wang1,*, Wangqi Du1,*, Chanqiong Zhang1, Yiling Song1, Yiqi Cai2, Danwei Cen1, Ledan Wang3, Yirong Xiong1, Pengfei Jiang1, Shanli Zhu1, Kong-Nan Zhao1, Lifang Zhang1
1Department of Microbiology and Immunology, Institute of molecular virology and immunology, Wenzhou Medical University, Wenzhou, China
2Department of General Surgery, First Affiliated Hospital, Wenzhou Medical University, Wenzhou, China
3Department of Obstetrics and Gynecology, Second Affiliated Hospital, Wenzhou Medical University, Wenzhou, China
*These authors have contributed equally to this work
Lifang Zhang, email: [email protected]
Keywords: cervical cancer, human papillomavirus, E7, affibody molecules, in vivo imaging
Received: April 20, 2016 Accepted: September 14, 2016 Published: September 21, 2016
Cervical cancer caused by infection with high-risk human papillomavirus remains to be the most deadly gynecologic malignancy worldwide. It is well documented that persistent expression of two oncogenes (E6/E7) plays the key roles in cervical cancer. Thus, in vivo detection of the oncoproteins is very important for the diagnosis of the cancer. Recently, affibody molecules have been demonstrated to be a powerful targeting probe for tumor–targeted imaging and diagnosis. In this study, four HPV16 E7-binding affibody molecules (Z HPV16 E7127, Z HPV16E7301, Z HPV16E7384 and Z HPV16E7745) were screened from a phage-displayed peptide library and used for molecular imaging in tumor-bearing mice. Biosensor binding analyses showed first that the four affibody molecules bound to HPV16 E7 with very high affinity and specificity. They co-localized with E7 protein only in two HPV16-positive cancer cells (SiHa and CaSki). Furthermore, affibody ZHPV16E7384 was conjugated with Dylight755 and used for in vivo tumor-imaging. Strongly high-contrast tumor retention of this affibody only occurred in HPV16-derived tumors of mice as early as 30 min post-injection, not in HPV-negative and HPV18-derived tumors. The accumulation of Dylight755-conjugated ZHPV16E7384 in tumor was achieved over a longer time period (24 h). The data here provide strong evidence that E7-specific affibody molecules have great potential used for molecular imaging and diagnosis of HPV-induced cancers.
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