Programmable protein arrays for immunoprofiling HPV-associated cancers

Radwa Ewaisha, Ian Meshay, Jack Resnik, Benjamin A. Katchman, Karen Anderson

    Research output: Contribution to journalArticle

    7 Citations (Scopus)

    Abstract

    Over 600 000 cancers each year are attributed to the human papillomavirus (HPV), including cervical, anogenital and oropharyngeal cancers (OPC). A key challenge in understanding HPV immunobiology is the diversity of oncogenic HPV types and the need for multiplexed display of HPV antigens to measure antibody responses. We have generated custom HPV protein microarrays displaying 98 proteins as C-terminal GST fusion proteins, representing eight antigens of two low-risk HPV types (HPV6 and 11) and ten oncogenic high-risk HPV types (HPV16, 18, 31, 33, 35, 39, 45, 51, 52 and 58). We demonstrate robust and reproducible protein expression of 96/98 of the antigens using a human cell lysate expression system. The target epitopes and specificities of four monoclonal antibodies were identified. Using sera from ten patients with newly diagnosed OPC and ten controls, we demonstrate specific IgG seroreactivity to HPV16 E1, E2, and E7 (a fold increase of 1.52, 2.19 and 1.35 in cases vs. controls, respectively, all p <0.005), confirming our prior data on an ELISA platform. We also detect HPV52 E7 Abs in serum from a patient with cervical cancer. The HPV protein array has potential for rapid identification of serologic responses to 12 HPV types.

    Original languageEnglish (US)
    Pages (from-to)1215-1224
    Number of pages10
    JournalProteomics
    Volume16
    Issue number8
    DOIs
    StatePublished - Apr 1 2016

    Fingerprint

    Protein Array Analysis
    Neoplasms
    Proteins
    Antigens
    Oropharyngeal Neoplasms
    Uterine Cervical Neoplasms
    Microarrays
    Epitopes
    Human papillomavirus 11
    Human papillomavirus 18
    Fusion reactions
    Immunoglobulin G
    Display devices
    Monoclonal Antibodies
    Cells
    Protein C
    Serum
    Antibodies
    Antibody Formation
    Enzyme-Linked Immunosorbent Assay

    Keywords

    • Antibodies
    • HPV
    • NAPPA
    • Protein microarrays
    • Serology

    ASJC Scopus subject areas

    • Molecular Biology
    • Biochemistry

    Cite this

    Programmable protein arrays for immunoprofiling HPV-associated cancers. / Ewaisha, Radwa; Meshay, Ian; Resnik, Jack; Katchman, Benjamin A.; Anderson, Karen.

    In: Proteomics, Vol. 16, No. 8, 01.04.2016, p. 1215-1224.

    Research output: Contribution to journalArticle

    Ewaisha, R, Meshay, I, Resnik, J, Katchman, BA & Anderson, K 2016, 'Programmable protein arrays for immunoprofiling HPV-associated cancers' Proteomics, vol. 16, no. 8, pp. 1215-1224. https://doi.org/10.1002/pmic.201500376
    Ewaisha, Radwa ; Meshay, Ian ; Resnik, Jack ; Katchman, Benjamin A. ; Anderson, Karen. / Programmable protein arrays for immunoprofiling HPV-associated cancers. In: Proteomics. 2016 ; Vol. 16, No. 8. pp. 1215-1224.
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