Format

Send to

Choose Destination
Oncotarget. 2018 Aug 24;9(66):32624-32641. doi: 10.18632/oncotarget.25976. eCollection 2018 Aug 24.

Quantitative screening of serum protein biomarkers by reverse phase protein arrays.

Kuang Z1,2, Huang R1,2, Yang Z3,4, Lv Z1, Chen X3,4, Xu F3,4, Yi YH1,5, Wu J6, Huang RP1,2,5.

Author information

1
RayBiotech Inc, Guangzhou, China.
2
RayBiotech Inc, Parkway Lane, Norcross, GA, USA.
3
Guangdong Provincial Academy of Chinese Medical Sciences, Guangzhou, China.
4
Guangdong Provincial Hospital of Chinese Medicine, Guangzhou, China.
5
South China Biochip Research Center, Guangzhou, China.
6
The Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China.

Abstract

Screening biomarkers in serum samples for different diseases has always been of great interest because it presents an early, reliable, and, most importantly, noninvasive means of diagnosis and prognosis. Reverse phase protein arrays (RPPAs) are a high-throughput platform that can measure single or limited sets of proteins from thousands of patients' samples in parallel. They have been widely used for detection of signaling molecules involved in diseases, especially cancers, and related regulation pathways in cell lysates. However, this approach has been difficult to adapt to serum samples. Previously, we developed a sensitive method called the enhanced protein array to quantitatively measure serum protein levels from large numbers of patient samples. Here, we further refine the technology on several fronts: 1. simplifying the experimental procedure; 2. optimizing multiple parameters to make the assay more robust, including the support matrix, signal reporting method, background control, and antibody validation; and 3. establishing a method for more accurate quantification. Using this technology, we quantitatively measured the expression levels of 10 proteins: alpha-fetoprotein (AFP), beta 2 microglobulin (B2M), Carcinoma Antigen 15-3(CA15-3), Carcinoembryonic antigen (CEA), golgi protein 73 (GP73), Growth differentiation factor 15 (GDF15), Human Epididymis Protein 4 (HE4), Insulin Like Growth Factor Binding Protein 2 (IGFBP2), osteopontin (OPN) and Beta-type platelet-derived growth factor receptor (PDGFRB) from serum samples of 132 hepatocellular carcinoma (HCC) patients and 78 healthy volunteers. We found that 6 protein expression levels are significantly increased in HCC patients. Statistical and bioinformatical analysis has revealed decent accuracy rates of individual proteins, ranging from 0.617 (B2M) to 0.908 (AFP) as diagnostic biomarkers to distinguish HCC from healthy controls. The combination of these 6 proteins as a specific HCC signature yielded a higher accuracy of 0.923 using linear discriminant analysis (LDA), logistic regression (LR), random forest (RF) and support vector machine (SVM) predictive model analyses. Our work reveals promise for using reverse phase protein arrays for biomarker discovery and validation in serum samples.

KEYWORDS:

biomarker screening; hepatocellular carcinoma; reverse phase protein array; serum protein

Conflict of interest statement

CONFLICTS OF INTEREST The authors declare no potential conflicts of interest.

Supplemental Content

Full text links

Icon for Impact Journals, LLC Icon for PubMed Central
Loading ...
Support Center