Development and characterization of an HPV18 detection kit using two novel HPV18 type-specific monoclonal antibodies

Zhang, Yao; He, Yan; Li, Ling; Liang, Shutian; Yan, Mei; Ren, Dongyan; Yang, Zengmin; Zhao, W X; Miao, Luyan; Zhang, Haijiang; Liu, Yongjiang

doi:10.1186/s13000-018-0727-7

Cited by 11 publications

(13 citation statements)

References 23 publications

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“…Therefore, detection of HPV E7 is essential for the diagnosis of cervical cancer in the early stage [21]. Recently, the HPV18 L1 ELISA detecting method and HPV16 E7-binding affibody molecules and nanobodies against the HPV16 E7 oncoprotein have been shown to have promising potential for the diagnosis of HPV-induced cancers [22][23][24]. The goal of this research is to establish a method with high sensitivity for quantitative detection of the HPV16 E7 oncoprotein in cervical cancer tissues.…”

Section: Discussionmentioning

confidence: 99%

Preparation, Characterization and Diagnostic Valuation of Two Novel Anti-HPV16 E7 Oncoprotein Monoclonal Antibodies

Dong

Zhang

et al. 2020

Viruses

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At present, the clinical detection method of human papillomavirus (HPV) is mainly based on the PCR method. However, this method can only be used to detect HPV DNA and HPV types, and cannot be used to accurately predict cervical cancer. HPV16 E7 is an oncoprotein selectively expressed in cervical cancers. In this study, we prepared an HPV16 E7-histidine (HIS) fusion oncoprotein by using a prokaryotic expression and gained several mouse anti-HPV16 E7-HIS fusion oncoprotein monoclonal antibodies (mAbs) by using hybridoma technology. Two mAbs, 69E2 (IgG2a) and 79A11 (IgM), were identified. Immunocytochemistry, immunofluorescence, immunohistochemistry, and Western blot were used to characterize the specificity of these mAbs. The sequences of the nucleotide bases and predicted amino acids of the 69E2 and 79A11 antibodies showed that they were novel antibodies. Indirect enzyme-linked immunosorbent assay (ELISA) with overlapping peptides, indirect competitive ELISA, and 3D structural modeling showed that mAbs 69E2 and 79A11 specifically bound to the three exposed peptides of the HPV16 E7 (HPV16 E7 49-66 , HPV16 E7 73-85 , and HPV16 E7 91-97 ). We used these two antibodies (79A11 as a capture antibody and 69E2 as a detection antibody) to establish a double-antibody sandwich ELISA based on a horseradish peroxidase (HRP)-labeled mAb and tetramethylbenzidine (TMB) detection system for quantitative detection of the HPV16 E7-HIS fusion oncoprotein, however, it was not ideal. Then we established a chemiluminescence immunoassay based on a labeled streptavidin-biotin (LSAB)-ELISA method and luminol detection system-this was sufficient for quantitative detection of the HPV16 E7-HIS fusion oncogenic protein in ng levels and was suitable for the detection of HPV16-positive cervical carcinoma tissues. Collectively, we obtained two novel mouse anti-HPV16 E7 oncoprotein mAbs and established an LSAB-lumino-dual-antibody sandwich ELISA method for the detection of the HPV16 E7-HIS fusion oncogenic protein, which might be a promising method for the diagnosis of HPV16-type cervical cancers in the early stage.Viruses 2020, 12, 333 3 of 23 carcinoma, and 5 cases of cervical cancer tissues derived from stage IIa HPV16-positive cervical cancers. All cancerous specimens were obtained by using the extensive hysterectomy, while normal specimens were obtained by using the panhysterectomy. A Mini Shaker MH-1 microplate oscillator (Kylin-Bell Lab Instruments, Haimen, Jiangsu, China), a 1510 MULTISKAN GO full-wavelength microplate reader (Thermo Fisher Scientific, Waltham, MA, USA), and an Infinite ® M200 PRO microplate reader (TECAN, Männedorf, Switzerland) were used for general experiments. Preparation of HPV16 E7-HIS Fusion OncoproteinAccording to the full-length HPV16 E7 gene, specific primers with 6 histidine (HIS) tags were designed. The sequences of primers F and R were as follows: F: 5 -GCCCATGGACCATCACCATC ACCATATGCATGGAGATACACCTAC-3 ; R: 5 -GCAAGCTTTTATGGTTTCTGYGAACAGATGG GGCACAC-3 . Using the total genome DNA of CaSki cells, which was pre-e...

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Section: Discussionmentioning

confidence: 99%

Preparation, Characterization and Diagnostic Valuation of Two Novel Anti-HPV16 E7 Oncoprotein Monoclonal Antibodies

Dong

Zhang

et al. 2020

Viruses

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“…In addition to oncogenic early proteins, many other kinds of anti-HPV antibodies, such as HPV L1 and L2, were also developed for diagnosis, evaluation of vaccines, or basal biomedical research. Two mAbs, specifically recognized as HPV18 L1 and VLPs, were used to develop an ELISA kit based on HRP-conjugated antibodies, and this method displayed good linearity, repeatability, and sensitivity for detecting HPV18 L1 pentamer and HPV18 VLP ( 31 ). Antibodies against HPV16 L1 can actively interact with the HPV18 L1, HPV31 L1, HPV45 L1, and HPV6 L1 ( 34 ).…”

Section: Immunological Methods Based On Hpv Proteins For Diagnosismentioning

confidence: 99%

“…According to DNA sequence analysis, HPVs have been divided into five genera-a, b, g, Nu, and Mu-each with different life cycle characteristics and related diseases (8)(9)(10). Epidemiological studies show that HPV types 16,18,31,33,35,39,45,51,52,56,58, and 59 are carcinogenic, and HPV68 are probably carcinogenic (6). Among them, HPV16 is the most prevalent worldwide and the major cause of HPV-associated cancers (11).…”

Section: Introductionmentioning

confidence: 99%

Immunodiagnosis and Immunotherapeutics Based on Human Papillomavirus for HPV-Induced Cancers

Dong

et al. 2021

Front. Immunol.

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Infection with human papillomavirus (HPV) is one of the main causes of malignant neoplasms, especially cervical, anogenital, and oropharyngeal cancers. Although we have developed preventive vaccines that can protect from HPV infection, there are still many new cases of HPV-related cancers worldwide. Early diagnosis and therapy are therefore important for the treatment of these diseases. As HPVs are the major contributors to these cancers, it is reasonable to develop reagents, kits, or devices to detect and eliminate HPVs for early diagnosis and therapeutics. Immunological methods are precise strategies that are promising for the accurate detection and blockade of HPVs. During the last decades, the mechanism of how HPVs induce neoplasms has been extensively elucidated, and several oncogenic HPV early proteins, including E5, E6, and E7, have been shown to be positively related to the oncogenesis and malignancy of HPV-induced cancers. These oncoproteins are promising biomarkers for diagnosis and as targets for the therapeutics of HPV-related cancers. Importantly, many specific monoclonal antibodies (mAbs), or newly designed antibody mimics, as well as new immunological kits, devices, and reagents have been developed for both the immunodiagnosis and immunotherapeutics of HPV-induced cancers. In the current review, we summarize the research progress in the immunodiagnosis and immunotherapeutics based on HPV for HPV-induced cancers. In particular, we depict the most promising serological methods for the detection of HPV infection and several therapeutical immunotherapeutics based on HPV, using immunological tools, including native mAbs, radio-labelled mAbs, affitoxins (affibody-linked toxins), intracellular single-chain antibodies (scFvs), nanobodies, therapeutical vaccines, and T-cell-based therapies. Our review aims to provide new clues for researchers to develop novel strategies and methods for the diagnosis and treatment of HPV-induced tumors.

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“…15,16,22 The use of PD-L1 as a predictive biomarker remains controversial given the lack of standardization among PD-L1 antibodies and detection assays, various PD-L1 expression cutoffs, and non-standardized test designs that make comparisons between trials difficult, 23,24 as discussed in detail elsewhere. 25 In addition, the unpredictability of responses to ICI therapies may be due in part to the heterogeneity and dynamics of PD-L1 expression by tumor cells and immune cells. 26,27 Recent reports indicate that PD-L1 expressed by both tumor cells and non-tumor host cells contributes to the efficacy of PD-1/PD-L1 checkpoint blockade therapies in preclinical models.…”

Section: Biomarkers Of Avelumab Activitymentioning

confidence: 99%

Product review: avelumab, an anti-PD-L1 antibody

Collins

Gulley

2018

Human Vaccines & Immunotherapeutics

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Although immunotherapies have been employed for many decades, immune checkpoint inhibitors have only recently entered the oncologic landscape. Avelumab is a fully human monoclonal antibody that blocks the interaction between PD-L1 on tumor cells and PD-1 on T cells, thereby inhibiting immunosuppression in the tumor microenvironment and reducing tumor growth. Most early clinical trials of avelumab as monotherapy and in combination regimens were part of the international JAVELIN clinical trial program, which included more than 7000 patients in more than 30 trials with at least 15 tumor types. Avelumab has been approved by the U.S. FDA for the treatment of metastatic Merkel cell carcinoma and metastatic urothelial carcinoma that has progressed during or following treatment with a platinum-based regimen. Its acceptable safety profile and ability to induce durable responses in otherwise deadly tumors provide the rationale for its use in other tumor types and in combination with other therapies.

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Development and characterization of an HPV18 detection kit using two novel HPV18 type-specific monoclonal antibodies

Cited by 11 publications

References 23 publications

Preparation, Characterization and Diagnostic Valuation of Two Novel Anti-HPV16 E7 Oncoprotein Monoclonal Antibodies

Preparation, Characterization and Diagnostic Valuation of Two Novel Anti-HPV16 E7 Oncoprotein Monoclonal Antibodies

Immunodiagnosis and Immunotherapeutics Based on Human Papillomavirus for HPV-Induced Cancers

Product review: avelumab, an anti-PD-L1 antibody

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