High-Throughput Protein Characterization Using Mass Spectrometric Immunoassay

Kiernan, Urban A.; Tubbs, Kemmons A.; Gruber, Karl; Nedelkov, Dobrin; Niederkofler, Eric E.; Williams, Peter W.; Nelson, Randall W.

doi:10.1006/abio.2001.5478

Cited by 72 publications

(62 citation statements)

References 43 publications

(28 reference statements)

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“…In our hands, sample throughput was higher for c-ELISA than for MS assays. In theory MS methods, however, can also be high throughput by automation (30,31 ). This technique requires robust protocols, because hepcidin readily sticks to laboratory plastics, especially with the use of small sample volumes.…”

Section: Discussionmentioning

confidence: 99%

Immunochemical and Mass-Spectrometry–Based Serum Hepcidin Assays for Iron Metabolism Disorders

Kroot¹,

Laarakkers²,

et al. 2010

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Section: Discussionmentioning

confidence: 99%

Immunochemical and Mass-Spectrometry–Based Serum Hepcidin Assays for Iron Metabolism Disorders

Kroot¹,

Laarakkers²,

et al. 2010

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“…Since its inception in 1995 by R.W. Nelson et al, 21 it has been applied to study a variety of proteins and peptides, [22][23][24][25][26][27][28][29][30][31][32][33][34][35][36] and has served as the foundation for the nascent field of Population Proteomics. [37][38][39][40][41][42] Roughly, MSIA can be envisioned as ultra high resolution, semiquantitative Western blotting in which protein variants are thoroughly resolved and their molecular masses determined to within less than 2 Da.…”

Section: Methodsmentioning

confidence: 99%

Glycosylation status of vitamin D binding protein in cancer patients

2009

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On the basis of the results of activity studies, previous reports have suggested that vitamin D binding protein (DBP) is significantly or even completely deglycosylated in cancer patients, eliminating the molecular precursor of the immunologically important Gc macrophage activating factor (GcMAF), a glycosidase-derived product of DBP. The purpose of this investigation was to directly determine the relative degree of O-linked trisaccharide glycosylation of serumderived DBP in human breast, colorectal, pancreatic, and prostate cancer patients. Results obtained by electrospray ionization-based mass spectrometric immunoassay showed that there was no significant depletion of DBP trisaccharide glycosylation in the 56 cancer patients examined relative to healthy controls. These results suggest that alternative hypotheses regarding the molecular and/or structural origins of GcMAF must be considered to explain the relative inability of cancer patient serum to activate macrophages.

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“…Its utility for delineating protein isoforms has been well documented (42)(43)(44)(45)(46)(47)(48)(49). In its simplest form, surface-immobilized ligands are utilized to affinity retrieve a protein of interest from a biological sample after which the protein (with or without the affinity ligand) is introduced in a mass spectrometer.…”

Section: Affinity-based Ms Methodsmentioning

confidence: 99%

Population Proteomics

Nedelkov

Kiernan

Niederkofler

et al. 2006

Molecular & Cellular Proteomics

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This review outlines the concept of population proteomics and its implication in the discovery and validation of cancer-specific protein modulations. Population proteomics is an applied subdiscipline of proteomics engaging in the investigation of human proteins across and within populations to define and better understand protein diversity. Population proteomics focuses on interrogation of specific proteins from large number of individuals, utilizing top-down, targeted affinity mass spectrometry approaches to probe protein modifications. Deglycosylation, sequence truncations, side-chain residue modifications, and other modifications have been reported for myriad of proteins, yet little is know about their incidence rate in the general population. Such information can be gathered via population proteomics and would greatly aid the biomarker discovery efforts. Discovery of novel protein modifications is also expected from such large scale population proteomics, expanding the protein knowledge database. In regard to cancer protein biomarkers, their validation via population proteomics-based approaches is advantageous as mass spectrometry detection is used both in the discovery and validation process, which is essential for the detection of those structurally modified protein biomarkers. Molecular & Cellular Proteomics 5: 1811-1818, 2006.In the past 5 years proteomics and its subdiscipline clinical proteomics (1-3) have taken a leading role in the discovery of new and improved cancer biomarkers. In its inception, clinical proteomics was an application of high end MS tools for evaluation of proteomic differences in the proteomes between healthy and cancer-bearing individuals. The tools were mainly based on the SELDI technology and utilization of wide specificity surfaces (e.g. hydrophobic or hydrophilic mass spectrometer targets) to bind groups of proteins from biological samples and generate distinct mass spectra containing hundreds of peaks (4, 5). These so-called proteomic patterns entered mainstream proteomics with the publication of a Lancet study in 2002 (6), which was quickly followed by a significant number of research studies (7-11) and reviews (12-15) describing proteomic patterns capable of discriminating between disease (e.g. ovarian cancer, prostate cancer, etc.) and healthy cohorts with relatively high sensitivity and specificity. However, critical assessment of those results quickly followed, outlining significant shortcomings and uncertainties in regard to the reproducibility of the findings, identity of the proteins behind the pattern peaks, and validation of the results (16 -23). Interlaboratory SELDI experiments performed recently alleviated some of the reproducibility concerns (24, 25). Furthermore, current research efforts have led to identification of several proteins behind the discriminating patterns peaks, including serum amyloid A (26), vitamin D-binding protein (27), and apolipoprotein A-II (28), which were identified as potential biomarkers for prostate cancer; haptoglobin (29), apolipopro...

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High-Throughput Protein Characterization Using Mass Spectrometric Immunoassay

Cited by 72 publications

References 43 publications

Immunochemical and Mass-Spectrometry–Based Serum Hepcidin Assays for Iron Metabolism Disorders

Immunochemical and Mass-Spectrometry–Based Serum Hepcidin Assays for Iron Metabolism Disorders

Glycosylation status of vitamin D binding protein in cancer patients

Population Proteomics

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