Detection of pancreatic cancer using antibody microarray‐based serum protein profiling

Abstract

The driving force behind oncoproteomics is to identify protein signatures that are associated with a particular malignancy. Here, we have used a recombinant scFv antibody microarray in an attempt to classify sera derived from pancreatic adenocarcinoma patients versus healthy subjects. Based on analysis of nonfractionated, directly labeled, whole human serum proteomes we have identified a protein signature based on 19 nonredundant analytes, that discriminates between cancer patients and healthy subjects. Furthermore, a potential protein signature, consisting of 21 protein analytes, could be defined that was shown to be associated with cancer patients having a life expectancy of <12 months. Taken together, the data suggest that antibody microarray analysis of complex proteomes will be a useful tool to define disease associated protein signatures.

Identification of Serum Biomarker Signatures Associated with Pancreatic Cancer

“…However, the current diagnostic tools are unable to perform accordingly and no high-performing biomarker(s) for PDACs are yet available (2, 24). We have recently described an affinity proteomic approach that allowed us to identify several serologic biomarker signatures, distinguishing other cancer indications from healthy controls, showing the strength of the technology platform (17,18,22,25). On the basis of the notion that immunoregulation is a particular phenomenon in PDACs (26,27), we thus undertook an affinity proteomics approach to harness the diagnostic power of the immune system to diagnose PDACs.…”

“…Patient cohort 2 was composed of 45 new patients, diagnosed with PDAC (n ¼ 25) or N (controls, n ¼ 20; for patient demographics, see ref. 17), as recently described (17). Patient cohorts 1 and 2 were from clinically independent sets of patients collected at different sites.…”

The Use of Antibodies in Small-Molecule Drug Discovery

“…11 Antibody arrays have recently been reported in multiple different formats, ranging from commercially available planar arrays, through qualitative multiplexed bead-based arrays and array-based surface plasmon resonance systems (which enable rapid, label-free, high-throughput analysis in low sample volume), to the greatly enhanced sensitivity of nanostructured immunoassays. [12][13][14][15] These technologies have been successfully applied in identifying global protein changes associated with a range of diseases including cancer [16][17][18][19] and neurodegenerative disorders. 20 The detailed technical aspects of these methodologies including some of the different technological characteristics have been reviewed previously by Borrebaeck and Wingren.…”

Generation of miniaturized planar ecombinant antibody arrays using a microcantilever-based printer

“…The specificity of these antibodies has previously been validated by using pure analytes; mixtures of pure analytes; well-characterized, standardized crude serum; and orthogonal methods, such as mass spectrometry (serum/tissue extract pull-down assays), ELISA, MesoScaleDiscovery (MSD) assay, immunohistochemistry, and/or cytometric bead assay; as well as by spiking and blocking experiments in crude sample formats (e.g. serum) [9,[40][41][42][43][44][45][46][47].…”