Catch and measure–mass spectrometry‐based immunoassays in biomarker research

Weiss, F. T.; Berg, Bart H. J. van den; Planatscher, Hannes; Pynn, Christopher J.; Joos, Thomas; Poetz, Oliver

doi:10.1016/j.bbapap.2013.09.010

Cited by 36 publications

(27 citation statements)

References 85 publications

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“…One of the initial aims of this study was to evaluate the capability to use the developed scFv antibodies for enrichment of the target proteins from a serum background followed by detection and quantification with the use of LC-SRM MS. Combining antibody enrichment with mass spectrometric readout has previously been demonstrated to enable detection of target proteins in complex sample formats such as serum or plasma [8][9][10] . More specifically and highly relevant to this study, it has been shown that recombinant scFv antibodies can be used in combination with LC-SRM MS for detection of proteins in a human serum background 11 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 while M1 in addition also binds the serum protein fibrinogen 28 .…”

Section: Discussionmentioning

confidence: 99%

Development of Phage-Based Antibody Fragment Reagents for Affinity Enrichment of Bacterial Immunoglobulin G Binding Proteins

et al. 2015

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Disease and death caused by bacterial infections are global health problems. Effective bacterial strategies are required to promote survival and proliferation within a human host, and it is important to explore how this adaption occurs. However, the detection and quantification of bacterial virulence factors in complex biological samples are technically demanding challenges. These can be addressed by combining targeted affinity enrichment of antibodies with the sensitivity of liquid chromatography-selected reaction monitoring mass spectrometry (LC-SRM MS). However, many virulence factors have evolved properties that make specific detection by conventional antibodies difficult. We here present an antibody format that is particularly well suited for detection and analysis of immunoglobulin G (IgG)-binding virulence factors. As proof of concept, we have generated single chain fragment variable (scFv) antibodies that specifically target the IgG-binding surface proteins M1 and H of Streptococcus pyogenes. The binding ability of the developed scFv is demonstrated against both recombinant soluble protein M1 and H as well as the intact surface proteins on a wild-type S. pyogenes strain. Additionally, the capacity of the developed scFv antibodies to enrich their target proteins from both simple and complex backgrounds, thereby allowing for detection and quantification with LC-SRM MS, was demonstrated. We have established a workflow that allows for affinity enrichment of bacterial virulence factors.

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Section: Discussionmentioning

confidence: 99%

Development of Phage-Based Antibody Fragment Reagents for Affinity Enrichment of Bacterial Immunoglobulin G Binding Proteins

et al. 2015

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“…Top-down MS-based approaches analyze intact proteins, without previous fragmentation to peptides [41,42]. These methods are better suited for proteoforms detection because they detect the mass of the intact proteoforms, and cover the putative modifications in the entire protein sequence.…”

Section: Introductionmentioning

confidence: 99%

Mass Spectrometric Immunoassays in Characterization of Clinically Significant Proteoforms

2016

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Proteins can exist as multiple proteoforms in vivo, as a result of alternative splicing and single-nucleotide polymorphisms (SNPs), as well as posttranslational processing. To address their clinical significance in a context of diagnostic information, proteoforms require a more in-depth analysis. Mass spectrometric immunoassays (MSIA) have been devised for studying structural diversity in human proteins. MSIA enables protein profiling in a simple and high-throughput manner, by combining the selectivity of targeted immunoassays, with the specificity of mass spectrometric detection. MSIA has been used for qualitative and quantitative analysis of single and multiple proteoforms, distinguishing between normal fluctuations and changes related to clinical conditions. This mini review offers an overview of the development and application of mass spectrometric immunoassays for clinical and population proteomics studies. Provided are examples of some recent developments, and also discussed are the trends and challenges in mass spectrometry-based immunoassays for the next-phase of clinical applications.

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“…Hence, those approaches cannot detect proteoforms without a priori knowledge of their existence. Top-down MS-based approaches are better suited for proteoforms detection because they detect the mass of the intact proteins, thus covering putative modifications in the entire protein sequence [9, 10]. …”

Section: Introductionmentioning

confidence: 99%

Development of multiplex mass spectrometric immunoassay for detection and quantification of apolipoproteins C-I, C-II, C-III and their proteoforms

Trenchevska

Schaab

Nelson

et al. 2015

Methods

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The impetus for discovery and evaluation of protein biomarkers has been accelerated by recent development of advanced technologies for rapid and broad proteome analyses. Mass spectrometry (MS)-based protein assays hold great potential for in vitro biomarker studies. Described here is the development of a multiplex Mass Spectrometric Immunoassay (MSIA) for quantification of apolipoprotein C-I (apoC-I), apolipoprotein C-II (apoC-II), apolipoprotein C-III (apoC-III) and their proteoforms. The multiplex MSIA assay was fast (~40 min) and high-throughput (96 samples at a time). The assay was applied to a small cohort of human plasma samples, revealing the existence of multiple proteoforms for each apolipoprotein C. The quantitative aspect of the assay enabled determination of the concentration for each proteoform individually. Low-abundance proteoforms, such as fucosylated apoC-III, were detected in less than 20% of the samples. The distribution of apoC-III proteoforms varied among samples with similar total apoC-III concentrations. The multiplex analysis of the three apolipoproteins C and their proteoforms using quantitative MSIA represents a significant step forward toward better understanding of their physiological roles in health and disease.

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Catch and measure–mass spectrometry‐based immunoassays in biomarker research

Cited by 36 publications

References 85 publications

Development of Phage-Based Antibody Fragment Reagents for Affinity Enrichment of Bacterial Immunoglobulin G Binding Proteins

Development of Phage-Based Antibody Fragment Reagents for Affinity Enrichment of Bacterial Immunoglobulin G Binding Proteins

Mass Spectrometric Immunoassays in Characterization of Clinically Significant Proteoforms

Development of multiplex mass spectrometric immunoassay for detection and quantification of apolipoproteins C-I, C-II, C-III and their proteoforms

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