Simultaneous Measurement of 92 Serum Protein Biomarkers for the Development of a Multiprotein Classifier for Ovarian Cancer Detection

Abstract

The best known ovarian cancer biomarker, CA125, is neither adequately sensitive nor specific for screening the general population. By using a combination of proteins for screening, it may be possible to increase the sensitivity and specificity over CA125 alone. In this study, we used Proseek Multiplex Oncology II plates to simultaneously measure the expression of 92 cancer-related proteins in serum using proximity extension assays. This technology combines the sensitivity of the PCR with the specificity of antibodybased detection methods, allowing multiplex biomarker detection and high-throughput quantification. We analyzed 1 mL of sera from each of 61 women with ovarian cancer and compared the values obtained with those from 88 age-matched healthy women. Principle component analysis and unsupervised hier-archical clustering separated the ovarian cancer patients from the healthy, with minimal misclassification. Data from the Proseek plates for CA125 levels exhibited a strong correlation with clinical values for CA125. We identified 52 proteins that differed significantly (P < 0.006) between ovarian cancer and healthy samples, several of which are novel serum biomarkers for ovarian cancer. In total, 40 proteins had an estimated area under the ROC curve of 0.70 or greater, suggesting their potential to serve as biomarkers for ovarian cancer. CA125 alone achieved a sensitivity of 93.4% at a specificity of 98%. By adding the Oncology II values for five proteins to CA125 in a multiprotein classifier, we increased the assay sensitivity to 98.4% at a specificity of 98%, thereby improving the sensitivity and specificity of CA125 alone.

A multiplex biomarker assay improves the diagnostic performance of HE4 and CA125 in ovarian tumor patients

“…their recent study on 92 biomarkers (Olink's Oncology II panel) reported lower levels of ITGAV in serum from ovarian cancer patients compared to healthy women [34].…”

“…In recent years, proximity extension assay (PEA) technology has been employed in the identification of novel protein biomarkers and biomarker combinations for early detection of ovarian cancer, with promising results [34,[48][49][50]. The addition of inflammatory and immunological biomarkers to CA125 and HE4 holds potential to increase sensitivity and specificity compared to the established algorithms.…”

Multi-biomarker panel signature as the key to diagnosis of ovarian cancer

“…Recently, Skubitz and group (Skubitz et al., 2019) has studied on multi-biomarkers that can differentiate between the serum samples of late stage high grade serous ovarian cancer and healthy control based on Proseek® Multiplex Oncology II plates. This plate can simultaneously measure the expression of 92 cancer related serum based protein biomarkers using proximity extension assays (PEA) (Assarsson et al., 2014; Thorsen et al., 2013; Chen et al., 2015).…”

“…In addition, to meet the minimum screening criteria of 99.6 % specificity, CA125 achieved a sensitivity of 85 %. Further, five biomarkers including FGFBP1, S100A4, EGF, ICOSLG, and MSLN, which were otherwise found at lower level with respect to control, on combination with CA125 enhances the sensitivity and specificity (Skubitz et al., 2019). This perhaps supports the inverse correlation study of additional biomarkers to primary markers (Pinsky and Zhu, 2011).…”

Evaluation of the potential of Pap test fluid and cervical swabs to serve as clinical diagnostic biospecimens for the detection of ovarian cancer by mass spectrometry-based proteomics

“…Future clinical applications may include ELISA tests using Pap test fluid or cervical swabs as the diagnostic biospecimen, or multiplexed proximity extension assays could be developed for use on Pap tests to sensitively quantify multiple proteins of interest [ 49 , 50 ]. Alternatively, SRM-based targeted proteomic assays are increasingly being used in the measurement of clinically significant proteins, allowing for cost effective, high throughput, sensitive, robust, multiplexed analysis and quantification [ 46 , 51 ].…”