A rapid test for the diagnosis of thrombotic thrombocytopenic purpura using surface enhanced laser desorption/ionization time‐of‐flight (SELDI‐TOF)‐mass spectrometry

Jin, Ming; Cataland, Spero R.; Bissell, M.G.; Wu, Haifeng

doi:10.1111/j.1538-7836.2006.01758.x

Cited by 72 publications

(76 citation statements)

References 24 publications

(41 reference statements)

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“…ADAMTS13 activity was determined using a surface enhanced laser desorption/ionization time-of-flight (SELDI-TOF) mass spectrometer (PCS4000), as reported previously. 10,19 All samples with ADAMTS13 activity less than 5% were retested using the modified protocol that determines ADAMTS13 activity precisely to levels as low as 0.5%. 19 ADAMTS13 autoantibody (IgG) was quantified using a commercially available ELISA kit from American Diagnostica Inc. (IMUBIND® ADAMTS13 Autoantibody ELISA), as described previously.…”

Section: Measurement Of Adamts13 Biomarkersmentioning

confidence: 99%

“…6,8 Several assays have been developed which rapidly detect ADAMTS13 activity and help to confirm a clinical suspicion of TTP. [10][11][12][13][14] An enzyme-linked immunosorbent assay (ELISA)-based test has also been used to measure ADAMTS13 antigen levels. 15,16 While ADAMTS13 activity measured at the onset of TTP is known to help to confirm the diagnosis, the clinical value of ADAMTS13 antigen measurements in the evaluation of TTP disease severity, responses to therapy, and clinical outcomes has not been well studied in a large cohort of TTP patients with longitudinal follow-up.…”

Section: Introductionmentioning

confidence: 99%

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ADAMTS13 activity and antigen during therapy and follow-up of patients with idiopathic thrombotic thrombocytopenic purpura: correlation with clinical outcome

Yang¹,

Jin²,

Lin³

et al. 2011

Haematologica

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BackgroundThe assay for ADAMTS13 activity helps clinicians to confirm the clinical diagnosis of idiopathic thrombotic thrombocytopenic purpura. The clinical value of testing for the antigen level of ADAMTS13 protein is, however, less clear. Design and MethodsIn this study, both ADAMTS13 antigen and activity levels were measured in 835 sequential samples from 40 consecutive patients who were followed for an average of 29 months throughout the course of acute episode plasma exchange treatment and clinical remission. ResultsDuring acute episodes, ADAMTS13 activity was severely deficient while ADAMTS13 antigen levels were more variable, ranging from severely deficient to as high as within the reference range. A severe depletion of ADAMTS13 antigen level during acute disease was, however, statistically associated with disease mortality (P=0.0322). For patients who achieved initial clinical responses, ADAMTS13 antigen levels appeared to be restored faster than ADAMTS13 activity to the normal range. Further analysis demonstrated that the ADAMTS13 antigen level at the time of initial clinical recovery was significantly higher in the patients who subsequently achieved a sustained clinical remission than in the group who soon after had an exacerbation (P=0.0187). ConclusionsOur data suggest that evaluation of ADAMTS13 antigen levels during the course of therapy and follow-up may offer additional useful information for the management of patients with thrombotic thrombocytopenic purpura.Key words: thrombotic thrombocytopenic purpura, ADAMTS13, prognosis, biomarker.Citation: Yang S, Jin M, Lin S, Cataland S, and Wu H. ADAMTS13 activity and antigen during therapy and follow-up of patients with idiopathic thrombotic thrombocytopenic purpura: correlation with clinical outcome Haematologica 2011;96(10)

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Section: Measurement Of Adamts13 Biomarkersmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

ADAMTS13 activity and antigen during therapy and follow-up of patients with idiopathic thrombotic thrombocytopenic purpura: correlation with clinical outcome

Yang¹,

Jin²,

Lin³

et al. 2011

Haematologica

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“…This demonstrates that the FCS assay is highly sensitive for rVWF-G1629E cleavage by ADAMTS13 at activities down to 0.5% of the physiological level. This feature could be exploited to improve the detection resolution of commonly used diagnostic assays, which lies between 2.5% and 5% (35,36). …”

Section: Cleavage Kinetics Of Rvwf-g1629ementioning

confidence: 99%

Mutation G1629E Increases von Willebrand Factor Cleavage via a Cooperative Destabilization Mechanism

Aponte-Santamaría

Lippok

Mittag

et al. 2017

Biophysical Journal

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The large multimeric glycoprotein von Willebrand Factor (VWF) plays a pivotal adhesive role during primary hemostasis. VWF is cleaved by the protease ADAMTS13 as a down-regulatory mechanism to prevent excessive VWF-mediated platelet aggregation. For each VWF monomer, the ADAMTS13 cleavage site is located deeply buried inside the VWF A2 domain. External forces in vivo or denaturants in vitro trigger the unfolding of this domain, thereby leaving the cleavage site solvent-exposed and ready for cleavage. Mutations in the VWF A2 domain, facilitating the cleavage process, cause a distinct form of von Willebrand disease (VWD), VWD type 2A. In particular, the VWD type 2A Gly1629Glu mutation drastically accelerates the proteolytic cleavage activity, even in the absence of forces or denaturants. However, the effect of this mutation has not yet been quantified, in terms of kinetics or thermodynamics, nor has the underlying molecular mechanism been revealed. In this study, we addressed these questions by using fluorescence correlation spectroscopy, molecular dynamics simulations, and free energy calculations. The measured enzyme kinetics revealed a 20-fold increase in the cleavage rate for the Gly1629Glu mutant compared with the wild-type VWF. Cleavage was found cooperative with a cooperativity coefficient n ¼ 2.3, suggesting that the mutant VWF gives access to multiple cleavage sites of the VWF multimer at the same time. According to our simulations and free energy calculations, the Gly1629Glu mutation causes structural perturbation in the A2 domain and thereby destabilizes the domain by~10 kJ/mol, promoting its unfolding. Taken together, the enhanced proteolytic activity of Gly1629Glu can be readily explained by an increased availability of the ADAMTS13 cleavage site through A2-domain-fold thermodynamic destabilization. Our study puts forward the Gly1629Glu mutant as a very efficient enzyme substrate for ADAMTS13 activity assays.

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“…The proteolytic processing of the reporter peptide Biotin-Abu-Ahx-WKPYDAADL-Ahx-HHHHHHt results in the accumulation of the anchor peptide AhxHHHHHHt (m/z 1055.1), and the respective peak intensities were used for quantification (15 ).…”

Section: Clinical Protease Profilingmentioning

confidence: 99%

“…This process results in the loss of MS signal intensity for transient fragments of reporter peptides when the incubation time is prolonged. Second, because serum is a very complex matrix, fractionation strategies, such as affinity chromatography (15 ), are essential to reduce unwanted ion suppression in MALDI-TOF MS analysis (16 ). Third, suitable reporter peptides with specific amino acid sequences that are selectively cleaved by tumor-associated proteases have to be identified in a systematic substrate screen (17,18 ).…”

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confidence: 99%

Endoprotease Profiling with Double-Tagged Peptide Substrates: A New Diagnostic Approach in Oncology

et al. 2010

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Background: The measurement of disease-related proteolytic activity in complex biological matrices like serum is of emerging interest to improve the diagnosis of malignant diseases. We developed a mass spectrometry (MS)-based functional proteomic profiling approach that tracks degradation of artificial endoprotease substrates in serum specimens. Methods: The synthetic reporter peptides that are cleaved by tumor-associated endopeptidases were systematically optimized with regard to flanking affinity tags, linkers, and stabilizing elements. Serum specimens were incubated with reporter peptides under standardized conditions and the peptides subsequently extracted with affinity chromatography before MS. In a pilot study an optimized reporter peptide with the cleavage motif WKPYDAADL was added to serum specimens from colorectal tumor patients (n = 50) and healthy controls (n = 50). This reporter peptide comprised a known cleavage site for the cysteine-endopeptidase “cancer procoagulant.” Results: Serial affinity chromatography using biotin- and 6xHis tags was superior to the single affinity enrichment using only 6xHis tags. Furthermore, protease-resistant stop elements ensured signal accumulation after prolonged incubation. In contrast, signals from reporter peptides without stop elements vanished completely after prolonged incubation owing to their total degradation. Reporter-peptide spiking showed good reproducibility, and the difference in proteolytic activity between serum specimens from cancer patients and controls was highly significant (P < 0.001). Conclusions: The introduction of a few structural key elements (affinity tags, linkers, d-amino acids) into synthetic reporter peptides increases the diagnostic sensitivity for MS-based protease profiling of serum specimens. This new approach might lead to functional MS-based protease profiling for improved disease classification.

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A rapid test for the diagnosis of thrombotic thrombocytopenic purpura using surface enhanced laser desorption/ionization time‐of‐flight (SELDI‐TOF)‐mass spectrometry

Cited by 72 publications

References 24 publications

ADAMTS13 activity and antigen during therapy and follow-up of patients with idiopathic thrombotic thrombocytopenic purpura: correlation with clinical outcome

ADAMTS13 activity and antigen during therapy and follow-up of patients with idiopathic thrombotic thrombocytopenic purpura: correlation with clinical outcome

Mutation G1629E Increases von Willebrand Factor Cleavage via a Cooperative Destabilization Mechanism

Endoprotease Profiling with Double-Tagged Peptide Substrates: A New Diagnostic Approach in Oncology

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