An experimental strategy for quantitative analysis of the humoral immune response to prostate cancer antigens using natural protein microarrays

Sara Forrester, Ji Qiu, Leslie Mangold, Alan Partin, David Misek, Brett Phinney, Douglas Whitten, Philip Andrews, Eleftherios Diamandis, Gilbert S. Omenn, Samir Hanash, Brian B. Haab

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

The identification of human tumor antigens has potential utility in the diagnosis and treatment of cancers. We demonstrate here a complete strategy to profile immunoreactivity and identify tumor antigens from proteins derived from tumor cell lines. Microarrays of proteins produced from 2-D LC fractionation of prostate tumor cell-line lysates were used to profile immunoreactivity in the sera of prostate cancer patients and control subjects. Cancer-associated immunoreactivity to distinct groups of chromatography fractions was present in about 50% of the patients, with greater immunoreactivity present in patients with non-organ-confined cancer than in patients with organ-confined cancer. We grouped the immunoreactive fractions by similarities in elution order and patterns of immunoreactivity to guide and interpret the MS analysis of selected fractions, which was used to identify the proteins that may be responsible for the immunoreactivity. As a complementary method to further characterize and validate the immunoreactivity of the proteins identified by mass spectrometry, we demonstrate the use of focused microarrays of recombinant proteins. Disease-associated immunoreactivity was confirmed for one of the identified proteins, human Kallikrein 11. These results demonstrate a practical approach to screening, identifying, and validating immunoreactive proteins that could be applied to diverse studies on humoral immune responses.

Original languageEnglish (US)
Pages (from-to)494-505
Number of pages12
JournalProteomics - Clinical Applications
Volume1
Issue number5
DOIs
StatePublished - May 2007
Externally publishedYes

Fingerprint

Protein Array Analysis
Microarrays
Humoral Immunity
Prostatic Neoplasms
Antigens
Neoplasm Antigens
Tumor Cell Line
Chemical analysis
Neoplasms
Proteins
Tumors
Forensic Anthropology
Cells
Recombinant Proteins
Chromatography
Prostate
Mass Spectrometry
Fractionation
Mass spectrometry
Screening

Keywords

  • Humoral immune response
  • Prostate cancer
  • Protein arrays

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

An experimental strategy for quantitative analysis of the humoral immune response to prostate cancer antigens using natural protein microarrays. / Forrester, Sara; Qiu, Ji; Mangold, Leslie; Partin, Alan; Misek, David; Phinney, Brett; Whitten, Douglas; Andrews, Philip; Diamandis, Eleftherios; Omenn, Gilbert S.; Hanash, Samir; Haab, Brian B.

In: Proteomics - Clinical Applications, Vol. 1, No. 5, 05.2007, p. 494-505.

Research output: Contribution to journalArticle

Forrester, S, Qiu, J, Mangold, L, Partin, A, Misek, D, Phinney, B, Whitten, D, Andrews, P, Diamandis, E, Omenn, GS, Hanash, S & Haab, BB 2007, 'An experimental strategy for quantitative analysis of the humoral immune response to prostate cancer antigens using natural protein microarrays', Proteomics - Clinical Applications, vol. 1, no. 5, pp. 494-505. https://doi.org/10.1002/prca.200600802
Forrester, Sara ; Qiu, Ji ; Mangold, Leslie ; Partin, Alan ; Misek, David ; Phinney, Brett ; Whitten, Douglas ; Andrews, Philip ; Diamandis, Eleftherios ; Omenn, Gilbert S. ; Hanash, Samir ; Haab, Brian B. / An experimental strategy for quantitative analysis of the humoral immune response to prostate cancer antigens using natural protein microarrays. In: Proteomics - Clinical Applications. 2007 ; Vol. 1, No. 5. pp. 494-505.
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