A novel multiplexed immunoassay identifies CEA, IL-8 and prolactin as prospective markers for Dukes’ stages A-D colorectal cancers

Abstract

BackgroundCurrent methods widely deployed for colorectal cancers (CRC) screening lack the necessary sensitivity and specificity required for population-based early disease detection. Cancer-specific protein biomarkers are thought to be produced either by the tumor itself or other tissues in response to the presence of cancers or associated conditions. Equally, known examples of cancer protein biomarkers (e.g., PSA, CA125, CA19-9, CEA, AFP) are frequently found in plasma at very low concentration (pg/mL-ng/mL). New sensitive and specific assays are therefore urgently required to detect the disease at an early stage when prognosis is good following surgical resection. This study was designed to meet the longstanding unmet clinical need for earlier CRC detection by measuring plasma candidate biomarkers of cancer onset and progression in a clinical stage-specific manner. EDTA plasma samples (1 μL) obtained from 75 patients with Dukes’ staged CRC or unaffected controls (age and sex matched with stringent inclusion/exclusion criteria) were assayed for expression of 92 human proteins employing the Proseek® Multiplex Oncology I proximity extension assay. An identical set of plasma samples were analyzed utilizing the Bio-Plex Pro™ human cytokine 27-plex immunoassay.ResultsSimilar quantitative expression patterns for 13 plasma antigens common to both platforms endorsed the potential efficacy of Proseek as an immune-based multiplex assay for proteomic biomarker research. Proseek found that expression of Carcinoembryonic Antigen (CEA), IL-8 and prolactin are significantly correlated with CRC stage.ConclusionsCEA, IL-8 and prolactin expression were found to identify between control (unaffected), non-malignant (Dukes’ A + B) and malignant (Dukes’ C + D) stages.Electronic supplementary materialThe online version of this article (doi:10.1186/s12014-015-9081-x) contains supplementary material, which is available to authorized users.

A multiplex platform for the identification of ovarian cancer biomarkers

“…The CA125 values that were obtained on the Proseek ® plates correlated with the clinical laboratory ELISA values, suggesting that the PEA technology used in the Proseek ® plates provides a comparable means of analysis of serum samples. Previous studies comparing ELISA values to PEA for biomarkers of colorectal cancer (CEA, IL-8, TIMP-1 and CA242) found correlations ranging from 0.73 to 0.98 [ 24 , 42 ]. In addition, Fredriksson et al [ 53 ] evaluated a multiplex proximity ligation assay (PLA), a precursor to PEA technology, for 20 biomarkers in plasma from 19 ovarian cancer patients of different histological subtypes and stages.…”

“…The reagents used in the Olink Proseek ® Multiplex 96-well plates are based on proximity extension assay technology [ 22 , 42 , 46 ], in which 96 oligonucleotide-labeled antibody pairs bind to their respective protein targets in the sample. When the oligo-tagged antibodies bind to the target protein in proximity to one another, a PCR reporter sequence is formed by DNA polymerization and subsequently detected and quantified using real-time PCR.…”

“…In particular, the panel contains CA125 and HE4, as well as 90 other cancer-related protein markers, including several that have been linked to ovarian cancer, such as ERBB2 [ 25 , 26 ], ERBB3 [ 27 ], ERBB4 [ 28 ], vascular endothelial growth factor receptor (VEGFR) 2 [ 29 ], midkine [ 30 – 33 ], kallikrein 6 [ 34 ], kallikrein 11 [ 35 ], folate receptor-alpha [ 36 ], interleukin-6 [ 37 – 40 ], and transforming growth factor-alpha [ 41 ]. Proseek ® Multiplex Oncology I plates were recently used to identify a panel of biomarkers for the detection of early stage colorectal cancers [ 42 , 43 ], while other studies have used Proseek ® plates for the discovery of bladder cancer biomarkers [ 44 ].…”

Proteomic profiling of human plasma for cancer biomarker discovery

“…Through protein microarray analysis, 43 of 8000 proteins were found to be differentially expressed in plasma samples from colorectal cancer patients, among which three proteins, PIM1, MAPKAPK3, and ACVR2B, were further validated by immunoblotting and immunohistochemistry . In a recent study, the Olink Proseek® Multiplex Oncology I proximity extension assay, which enables high‐throughput, multiplex immunoassays that measure 92 proteins across 96 samples simultaneously using only 1 μL of serum or plasma was used to analyze the plasma of CRC patients and control samples, showing CEA, IL‐8, and prolactin as three potential biomarkers capable of predicting different stages of CRC .…”

Mucin 16 and kallikrein 13 as potential prognostic factors in colon cancer: Results of an oncological 92-multiplex immunoassay

“…3 Other tumor markers, such as IL-8, prolactin, and carbohydrate antigen 19-9 (CA19-9), are comparable to CEA, but fall short in terms of adding value to prognostics. 4,5 However, an elevated CA19-9 level in CRC patients with lymph node invasion serves as an independent prognostic marker and may carry further prognostic value alongside an elevated CEA level. 5,6 We continue to need new and accurate blood-based biomarkers.…”