Quantitative analysis of Tenecteplase in rat plasma samples using LC–MS/MS as an alternative for ELISA

Buscher, B.A.P.; Gerritsen, H.W.; Schöll, I. van; Cnubben, N.H.P.; Brull, L

doi:10.1016/j.jchromb.2006.12.053

Cited by 21 publications

(13 citation statements)

References 14 publications

(13 reference statements)

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“…The concentration of GnHCl was reduced to 0.9 M prior to the addition of trypsin, as recommended by Promega40 and in accordance with values found in the literature 19. However, in this study we found that GnHCl-containing digests consistently showed lower efficiencies of digestion, resulting in lower peak area ratios in the MRM analysis than with the other methods.…”

Section: Resultssupporting

confidence: 88%

“…Guanidine HCl Protocol. The GnHCl digestion protocol of Buscher et al 19 in which 1.0 M GnHCl was used was modified in accordance with the instructions in the Promega trypsin product insert. 20 The GnHCl concentration was kept below 1.0 M during tryptic digestion in order to maintain the activity of the trypsin in the presence of this strong chaotropic agent.…”

Section: Stable-isotope Labeled Internal Standard (Sis) Peptidesmentioning

confidence: 99%

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A Quantitative Study of the Effects of Chaotropic Agents, Surfactants, and Solvents on the Digestion Efficiency of Human Plasma Proteins by Trypsin

et al. 2010

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Plasma biomarkers studies are based on the differential expression of proteins between different treatment groups or between diseased and control populations. Most mass spectrometry-based methods of protein quantitation, however, are based on the detection and quantitation of peptides, not intact proteins. For peptide-based protein quantitation to be accurate, the digestion protocols used in proteomic analyses must be both efficient and reproducible. There have been very few studies, however, where plasma denaturation/digestion protocols have been compared using absolute quantitation methods. In this paper, 14 combinations of heat, solvent [acetonitrile, methanol, trifluoroethanol], chaotropic agents [guanidine hydrochloride, urea], and surfactants [sodium dodecyl sulfate (SDS) and sodium deoxycholate (DOC)] were compared with respect to their effectiveness in improving subsequent tryptic digestion. These digestion protocols were evaluated by quantitating the production of proteotypic tryptic peptides from 45 moderate- to high-abundance plasma proteins, using tandem mass spectrometry in multiple reaction monitoring mode, with a mixture of stable-isotope labeled analogues of these proteotypic peptides as internal standards. When the digestion efficiencies of these 14 methods were compared, we found that both of the surfactants (SDS and DOC) produced an increase in the overall yield of tryptic peptides from these 45 proteins, when compared to the more commonly used urea protocol. SDS, however, can be a serious interference for subsequent mass spectrometry. DOC, on the other hand, can be easily removed from the samples by acid precipitation. Examining the results of a reproducibility study, done with 5 replicate digestions, DOC and SDS with a 9 h digestion time produced the highest average digestion efficiencies (~80%), with the highest average reproducibility (<5% error, defined as the relative deviation from the mean value). However, because of potential interferences resulting from the use of SDS, we recommend DOC with a 9 h digestion procedure as the optimum protocol.

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Section: Resultssupporting

confidence: 88%

Section: Stable-isotope Labeled Internal Standard (Sis) Peptidesmentioning

confidence: 99%

A Quantitative Study of the Effects of Chaotropic Agents, Surfactants, and Solvents on the Digestion Efficiency of Human Plasma Proteins by Trypsin

et al. 2010

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“…[473] The detection of proteins has been instrumental in medicine, and has been commonly done now for decades through western blots and ELISAs, [474,475] as well as some more comprehensive approaches used to validate ELISA results (e.g., liquid chromatography-mass spectrometry, LC-MS). [476,477] Most biosensors involved in protein detection still function on the traditional antibody/antigen or antibody/antibody interactions that are frequently used in biological detection. For example, biosensors can be designed to detect antibodies to a particular infectious agent, confirming whether an individual has previously been infected, mounting an immune response.…”

Section: Clinical Applications Of Nanomaterials In Diagnosticsmentioning

confidence: 99%

Advances in Biosensors and Diagnostic Technologies Using Nanostructures and Nanomaterials

Welch

Powell

Clevinger

et al. 2021

Adv Funct Materials

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Nanoscale materials have unique properties that make them especially useful for biomedical diagnostic applications. Recent developments in nanoengineering have resulted in increasing use of nanostructures in biosensors. Various types of 0D, 1D, 2D, and 3D nanostructures have been used to improve biosensor sensitivity, selectivity, limit of detection, and time to result, among other metrics. These nanostructures have been integrated into electrochemical, optical, and other biosensors for this purpose. Here, the most recent advances in the use of nanostructured materials in biosensors are described. This includes a discussion of nanoparticles, nanorods, nanofibers, nanopillars, nanowires, nanosheets, indented nanopatterns (nanoholes and nanoslits), nanogaps, nanochannels, nanopores, nanofunctionalized surfaces, and complex hierarchical structures and their unique advantages and applications in biosensors. Clinical applications of these nanobiosensors are also highlighted along with a discussion of the future directions for these materials in diagnostics.

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“…However, there were successful applications with the 'no IS' approach. Buscher et al shared their experience of 'no IS' quantification of protein and semi-validated the method, which was then applied successfully in a rat preclinical study [77]. Although it is not widely published, the 'no IS' approach is often used in our laboratory since we support the quantification of proteins mostly still in the discovery stage.…”

Section: No Ismentioning

confidence: 99%

Application and Challenges in Using LC–MS Assays for Absolute Quantitative Analysis of Therapeutic Proteins in Drug Discovery

et al. 2014

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As more protein therapeutics enter the drug-discovery pipeline, the traditional ligand-binding assay (LBA) faces additional challenges to meet the rapid and diverse bioanalytical needs in the early drug-discovery stage. The high specificity and sensitivity afforded by LC-MS, along with its rapid method development, is proving invaluable for the analysis of protein therapeutics in support of drug discovery. LC-MS not only serves as a quantitative tool to complement LBA in drug discovery, it also provides structural details at a molecular level, which are used to address issues that cannot be resolved using LBA alone. This review will describe the key benefits and applications, as well as the techniques and challenges for applying LC-MS to support protein quantification in drug discovery.

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Quantitative analysis of Tenecteplase in rat plasma samples using LC–MS/MS as an alternative for ELISA

Cited by 21 publications

References 14 publications

A Quantitative Study of the Effects of Chaotropic Agents, Surfactants, and Solvents on the Digestion Efficiency of Human Plasma Proteins by Trypsin

A Quantitative Study of the Effects of Chaotropic Agents, Surfactants, and Solvents on the Digestion Efficiency of Human Plasma Proteins by Trypsin

Advances in Biosensors and Diagnostic Technologies Using Nanostructures and Nanomaterials

Application and Challenges in Using LC–MS Assays for Absolute Quantitative Analysis of Therapeutic Proteins in Drug Discovery

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