Programmable protein arrays for immunoprofiling HPV-associated cancers

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

Research output: Contribution to journalArticle

8 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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