Antibody-based enrichment of peptides on magnetic beads for mass-spectrometry-based quantification of serum biomarkers

Whiteaker, Jeffrey R.; Zhao, Lei; Zhang, Heidi Y.; Li, Feng; Piening, Brian D.; Anderson, Leigh; Paulovich, Amanda G.

doi:10.1016/j.ab.2006.12.023

Cited by 256 publications

(211 citation statements)

References 38 publications

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“…23,24 However, these techniques are not suitable for the assignment of values to reference materials. A reference measurement procedure must be critically investigated as to all potential sources of error, which must be identified, evaluated, and mitigated to the greatest extent possible.…”

Section: Discussionmentioning

confidence: 99%

Reference Measurement Procedure Development for C-Reactive Protein in Human Serum

Kilpatrick

Bunk

2009

Anal. Chem.

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This paper describes the development of a reference measurement procedure to quantify human C-reactive protein (CRP) in serum using affinity techniques prior to tryptic digestion and liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the certification of reference materials in clinically relevant ranges. The absence of a suitable internal standard for the CRP measurement, necessary to eliminate potential measurement bias in both the affinity purification and trypsin digestion steps, was addressed using the method of standard addition. The standard addition quantification approach was combined with affinity purification, using an anti-CRP monoclonal antibody conjugated to polystyrene beads, trypsin digestion of the purified protein, and LC-MS/MS analysis of CRP tryptic peptides. The effectiveness of intact protein affinity purification was evaluated through the measurement of CRP in several serum-based CRP control materials, yielding levels that were comparable to their expected mean concentration values. Quantitative results were confirmed with an external calibration approach. This study demonstrates the feasibility of affinity purification with LC-MS/MS for the reference measurement procedure development of low abundance serum protein analytes.Reference measurement procedures are used for a variety of purposes including (1) the assessment of the measurement quality of routine measurement procedures, (2) the value assignment of high-order calibration solutions (the so-called "master calibrators") for routine assays, and (3) the value assignment of certified reference materials. 1,2 In these roles, reference measurement procedures are considered "higher-order" measurement procedures, as they aim to achieve a higher level of accuracy and precision than the routine assay for the analyte being measured, producing a value with a low uncertainty of defined magnitude. In order for a reference measurement procedure to be fit for this purpose, it must be thoroughly evaluated to identify sources of bias and assess their magnitude. [3][4][5] Minimizing or eliminating bias is necessary to measure the most accurate, true concentration of the analyte. Because of the need for high levels of accuracy and precision, reference measurement procedures are typically low-throughput, labor intensive, and often costly to perform, in sharp contrast to what is desirable for routine measurements.Reference measurement procedures are key elements in the establishment of measurement standardization and metrological traceability in clinical chemistry. For more than 30 years, isotope dilution mass spectrometry (IDMS) has been one of the analytical techniques used frequently in reference measurement procedures, particularly for small molecule and inorganic analytes of clinical importance. 6 Despite this, few reference measurement procedures exist for clinically relevant proteins. This is one reason why so many clinical protein analytes are measured in arbitrary International Units (IUs); without a reference measurement procedure to...

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Section: Discussionmentioning

confidence: 99%

Reference Measurement Procedure Development for C-Reactive Protein in Human Serum

Kilpatrick

Bunk

2009

Anal. Chem.

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“…Both relative and absolute quantification information about the protein of interest is then gathered [49]. This method was further optimized for greater precision and accuracy by Whiteaker et al, reporting a physiologically relevant sensitivity in the ng/mL range [50]. Whiteaker et al also utilized the a combination of antibody and MS approaches to confirm global proteomics results from mouse model of breast cancer while characterizing and transferring a couple of the proteins identified previously to a plasma biomarker [38].…”

Section: Additional Targeted Proteomics Methodsmentioning

confidence: 99%

Current affairs in quantitative targeted proteomics: multiple reaction monitoring-mass spectrometry

Yocum¹,

Chinnaiyan²

2009

Briefings in Functional Genomics and Proteomics

109

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Quantitative targeted proteomics has recently taken front stage in the proteomics community. Centered on multiple reaction monitoring^mass spectrometry (MRM^MS) methodologies, quantitative targeted proteomics is being used in the verification of global proteomics data, the discovery of lower abundance proteins, protein post-translational modifications, discrimination of select highly homologous protein isoforms and as the final step in biomarker discovery. An older methodology utilized with small molecule analysis, the proteomics community is making great technological strides to develop MRM^MS as the next method to address previously challenging issues in global proteomics experimentation, namely dynamic range, identification of post-translational modifications, sensitivity and selectivity of measurement which will undoubtedly further biomedical knowledge.This brief review will provide a general introduction of MRM^MS and highlight its novel application for targeted quantitative proteomic experimentations.

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“…One family of nanomaterials favoured with purification strategies is Superparamagnetic Particles, SPPs. These particles allow for the removal of specific analytes from complex sample matrices using nothing more complicated than a hand held magnet 14,[20][21][22][23] and the use of SPPs has become increasingly common. When they are incorporated into fluidic devices they can be used to continuously sort cells and DNA from liquids 24 , and are integrated into a variety of detection platforms 14,24,25 .…”

Section: Introductionmentioning

confidence: 99%

Particle-by-Particle Charge Analysis of DNA-Modified Nanoparticles Using Tunable Resistive Pulse Sensing

2016

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Resistive pulse sensors, RPS, are allowing the transport mechanism of molecules, proteins and even nanoparticles to be characterised as they traverse pores. Previous work using RPS has shown that the size, concentration and zeta potential of the analyte can be measured. Here we use tunable resistive pulse sensing (TRPS) which utilises a tunable pore to monitor the translocation times of nanoparticles with DNA modified surfaces. We start by demonstrating that the translocation times of particles can be used to infer the zeta potential of known standards and then apply the method to measure the change in zeta potential of DNA modified particles. By measuring the translocation times of DNA modified nanoparticles as a function of packing density, length, structure, and hybridisation time, we observe a clear difference in zeta potential using both mean values, and population distributions as a function of the DNA structure. We demonstrate the ability to resolve the signals for ssDNA, dsDNA, small changes in base length for nucleotides between 15-40 bases long and even the discrimination between partial and fully complementary target sequences. Such a method has potential and applications in sensors for the monitoring of nanoparticles in both medical and environmental samples.

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Antibody-based enrichment of peptides on magnetic beads for mass-spectrometry-based quantification of serum biomarkers

Cited by 256 publications

References 38 publications

Reference Measurement Procedure Development for C-Reactive Protein in Human Serum

Reference Measurement Procedure Development for C-Reactive Protein in Human Serum

Current affairs in quantitative targeted proteomics: multiple reaction monitoring-mass spectrometry

Particle-by-Particle Charge Analysis of DNA-Modified Nanoparticles Using Tunable Resistive Pulse Sensing

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