title: Generation of affibody molecules specific for HPV16 E7 recognition.
abstract: 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.
authors.affiliation: Department of Microbiology and Immunology, Institute of molecular virology and immunology, Wenzhou Medical University, Wenzhou, China.
authors.forename: Xiangyang
authors.initials: X
authors.lastname: Xue
authors.affiliation: Department of Microbiology and Immunology, Institute of molecular virology and immunology, Wenzhou Medical University, Wenzhou, China.
authors.forename: Bingbing
authors.initials: B
authors.lastname: Wang
authors.affiliation: Department of Microbiology and Immunology, Institute of molecular virology and immunology, Wenzhou Medical University, Wenzhou, China.
authors.forename: Wangqi
authors.initials: W
authors.lastname: Du
authors.affiliation: Department of Microbiology and Immunology, Institute of molecular virology and immunology, Wenzhou Medical University, Wenzhou, China.
authors.forename: Chanqiong
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Department of Microbiology and Immunology, Institute of molecular virology and immunology, Wenzhou Medical University, Wenzhou, China.
authors.forename: Yiling
authors.initials: Y
authors.lastname: Song
authors.affiliation: Department of General Surgery, First Affiliated Hospital, Wenzhou Medical University, Wenzhou, China.
authors.forename: Yiqi
authors.initials: Y
authors.lastname: Cai
authors.affiliation: Department of Microbiology and Immunology, Institute of molecular virology and immunology, Wenzhou Medical University, Wenzhou, China.
authors.forename: Danwei
authors.initials: D
authors.lastname: Cen
authors.affiliation: Department of Obstetrics and Gynecology, Second Affiliated Hospital, Wenzhou Medical University, Wenzhou, China.
authors.forename: Ledan
authors.initials: L
authors.lastname: Wang
authors.affiliation: Department of Microbiology and Immunology, Institute of molecular virology and immunology, Wenzhou Medical University, Wenzhou, China.
authors.forename: Yirong
authors.initials: Y
authors.lastname: Xiong
authors.affiliation: Department of Microbiology and Immunology, Institute of molecular virology and immunology, Wenzhou Medical University, Wenzhou, China.
authors.forename: Pengfei
authors.initials: P
authors.lastname: Jiang
authors.affiliation: Department of Microbiology and Immunology, Institute of molecular virology and immunology, Wenzhou Medical University, Wenzhou, China.
authors.forename: Shanli
authors.initials: S
authors.lastname: Zhu
authors.affiliation: Department of Microbiology and Immunology, Institute of molecular virology and immunology, Wenzhou Medical University, Wenzhou, China.
authors.forename: Kong-Nan
authors.initials: KN
authors.lastname: Zhao
authors.affiliation: Department of Microbiology and Immunology, Institute of molecular virology and immunology, Wenzhou Medical University, Wenzhou, China.
authors.forename: Lifang
authors.initials: L
authors.lastname: Zhang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D000918:Antibody Specificity / Q000276:immunology*; D045744:Cell Line, Tumor; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D051379:Mice; D057054:Molecular Imaging / Q000379:methods; D061848:Optical Imaging / Q000379:methods; D050725:Papillomavirus E7 Proteins / Q000276:immunology* / Q000378:metabolism; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D002583:Uterine Cervical Neoplasms / Q000000981:diagnostic imaging / Q000209:etiology / Q000473:pathology
chemical_list: D000906:Antibodies; D050725:Papillomavirus E7 Proteins; D011993:Recombinant Fusion Proteins; C059731:oncogene protein E7, Human papillomavirus type 16
keywords: E7; affibody molecules; cervical cancer; human papillomavirus; in vivo imaging