Validation of reverse phase protein array for practical screening of potential biomarkers in serum and plasma: Accurate detection of CA19-9 levels in pancreatic cancer

Tobias Grote, Doris R. Siwak, Herbert A. Fritsche, Corwin Joy, Gordon Mills, Diane Simeone, David C. Whitcomb, Craig D. Logsdon

Research output: Contribution to journalArticle

57 Citations (Scopus)

Abstract

The current study analyzed reverse phase protein arrays (RPPA) as a means to experimentally validate biomarkers in blood samples. One microliter samples of sera (n = 71), and plasma (n = 78) were serially diluted and printed on NC-coated slides. CA19-9 levels from RPPA results were compared with identical patient samples as measured by ELISA. There was a strong correlation between RPPA and ELISA (r = 0.87) as determined by scatter plots. Sample reproducibility of CA19-9 levels was excellent (interslide correlation r = 0.88; intraslide correlation r = 0.83). The ability of RPPA to accurately distinguish CA19-9 levels between cancer and noncancer samples were determined using receiver operating characteristic curves and compared with ELISA. The AUC for RPPA and ELISA was comparable (0.87 and 0.86, respectively). When the mean CA19-9 levels of normal samples was used as a cutoff for RPPA and compared with the standard clinical ELISA cutoff, comparable specificities (71% for both) were observed. Notably, RPPA samples normalized to albumin showed increased sensitivity compared to ELISA (90% vs. 75%). As RPPA is a high-throughput method that shows results comparable to that of ELISA, we propose that RPPA is a viable technique for rapid experimental screening and validation of candidate biomarkers in blood samples.

Original languageEnglish (US)
Pages (from-to)3051-3060
Number of pages10
JournalProteomics
Volume8
Issue number15
DOIs
StatePublished - Aug 1 2008
Externally publishedYes

Fingerprint

Protein Array Analysis
Biomarkers
Pancreatic Neoplasms
Screening
Plasmas
Enzyme-Linked Immunosorbent Assay
Serum
Proteins
Blood
ROC Curve
Area Under Curve
Albumins
Throughput

Keywords

  • Biomarker
  • CA19-9
  • Pancreatic cancer
  • Reverse phase protein array

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Validation of reverse phase protein array for practical screening of potential biomarkers in serum and plasma : Accurate detection of CA19-9 levels in pancreatic cancer. / Grote, Tobias; Siwak, Doris R.; Fritsche, Herbert A.; Joy, Corwin; Mills, Gordon; Simeone, Diane; Whitcomb, David C.; Logsdon, Craig D.

In: Proteomics, Vol. 8, No. 15, 01.08.2008, p. 3051-3060.

Research output: Contribution to journalArticle

Grote, Tobias ; Siwak, Doris R. ; Fritsche, Herbert A. ; Joy, Corwin ; Mills, Gordon ; Simeone, Diane ; Whitcomb, David C. ; Logsdon, Craig D. / Validation of reverse phase protein array for practical screening of potential biomarkers in serum and plasma : Accurate detection of CA19-9 levels in pancreatic cancer. In: Proteomics. 2008 ; Vol. 8, No. 15. pp. 3051-3060.
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