Amine-reactive isobaric tagging reagents: Requirements for absolute quantification of proteins and peptides

2008 ◽  
Vol 379 (2) ◽  
pp. 164-169 ◽  
Author(s):  
M. Quaglia ◽  
C. Pritchard ◽  
Z. Hall ◽  
G. O’Connor
Keyword(s):  
Absolute Quantification ◽  
Isobaric Tagging ◽  
Proteins And Peptides

Quantitative proteomic analyses of Schistosoma japonicum in response to artesunate

2015 ◽  
Vol 11 (5) ◽  
pp. 1400-1409 ◽  
Author(s):  
QingMing Kong ◽  
QunBo Tong ◽  
Di Lou ◽  
JianZu Ding ◽  
Bin Zheng ◽  
...  
Keyword(s):  
Schistosoma Japonicum ◽  
Absolute Quantification ◽  
Proteomic Analyses ◽  
Isobaric Tagging

We applied isobaric tagging reagents for relative and absolute quantification analyses to investigate the effect of artesunate on the proteome ofSchistosoma japonicumin susceptible mice.

scholarly journals Using bioinformatics for the identification of key peptides to engineer dopamine neurons. Towards a therapy for Parkinson’s disease.

2018 ◽  
Author(s):  
William D Mitchell ◽  
Rowan P Orme ◽  
Sarah R Hart ◽  
Rosemary A Fricker
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Absolute Quantification ◽  
Dopamine Neurons ◽  
Neuron Development ◽  
Expression Levels ◽  
Relative Expression ◽  
Online Databases ◽  
Isobaric Tagging

Parkinson’s disease is a widespread condition caused by degeneration of dopamine neurons in the midbrain. A number of proteins are known to be important to signalling mechanisms present in the midbrain during natural dopamine neuron development, and may be utilised to better produce dopamine neurons in vitro. Relative expression levels of proteins were obtained from substantia nigra tissue of rats from embryonic days E11 through E14 using isobaric tagging for relative and absolute quantification. This project analysed the dataset obtained, with an emphasis on relative expression levels of proteins across the four-day period. Bioinformatics searching of online databases reduced the dataset from 3325 proteins to a shortlist of five worthy of further investigation. It is hoped that the proteins identified using these techniques will help to refine protocols for the production of dopamine neurons in vitro.

scholarly journals Fully Traceable Absolute Protein Quantification of Somatropin That Allows Independent Comparison of Somatropin Standards

2009 ◽  
Vol 55 (11) ◽  
pp. 1984-1990 ◽  
Author(s):  
Caroline Pritchard ◽  
Milena Quaglia ◽  
Chris Mussell ◽  
William I Burkitt ◽  
Helen Parkes ◽  
...  
Keyword(s):  
Clinical Chemistry ◽  
Recombinant Human Growth Hormone ◽  
Isotope Dilution Mass Spectrometry ◽  
Standard Addition ◽  
Absolute Quantification ◽  
Protein Quantification ◽  
Affinity Tag ◽  
Digestion Method ◽  
Amount Of Substance ◽  
Isobaric Tagging

Abstract Background: Measurement traceability in clinical chemistry is required to standardize clinical results irrespective of the measurement procedure and laboratory. The traceability of many protein substances is maintained by reference to the first standard produced, which may no longer exist, with values assigned by consensus. Independent methods that provide traceability to the Système d’Unité International for all relevant properties of a protein standard could remove reliance on the original standard preparations. Methods: We developed a method based on the traceable quantification of tryptic peptides released from the protein by isotope dilution mass spectrometry to compare 2 standard preparations of somatropin (recombinant human growth hormone), WHO 98/574 and Ph.Eur.CRS S0947000. Relative quantification using isotope-coded affinity tagging, isobaric tagging for relative and absolute quantification, and standard additions were also performed to validate the digestion method used and to determine whether any modifications were present. Results: The total somatropin content in both materials was determined and an uncertainty estimation undertaken [WHO 2.19 ± 0.21) mg/vial, European Pharmacopeia 2.06 ± 0.21 mg/vial]. Each uncertainty in this paper is a fully estimated uncertainty, with 95% CI (k = 2). Isotope coded affinity tag and standard addition results fully validated the robustness of the digestion method used. In addition, iTRAQ (isobaric tagging for relative and absolute quantification analysis) identified 2 modifications, neither of which impacted the quantification. Conclusions: An independent method that does not rely on a preexisting protein standard has been developed and validated for the traceable value-assignment of total somatropin. The methods reported here address the amount of substance (mass fraction) of the standard materials but address neither biological activity nor other characteristics that may be important in assessing suitability for use as a calibrator.

scholarly journals Using bioinformatics for the identification of key peptides to engineer dopamine neurons. Towards a therapy for Parkinson’s disease.

2018 ◽  
Author(s):  
William D Mitchell ◽  
Rowan P Orme ◽  
Sarah R Hart ◽  
Rosemary A Fricker
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Absolute Quantification ◽  
Dopamine Neurons ◽  
Neuron Development ◽  
Expression Levels ◽  
Relative Expression ◽  
Online Databases ◽  
Isobaric Tagging

Parkinson’s disease is a widespread condition caused by degeneration of dopamine neurons in the midbrain. A number of proteins are known to be important to signalling mechanisms present in the midbrain during natural dopamine neuron development, and may be utilised to better produce dopamine neurons in vitro. Relative expression levels of proteins were obtained from substantia nigra tissue of rats from embryonic days E11 through E14 using isobaric tagging for relative and absolute quantification. This project analysed the dataset obtained, with an emphasis on relative expression levels of proteins across the four-day period. Bioinformatics searching of online databases reduced the dataset from 3325 proteins to a shortlist of five worthy of further investigation. It is hoped that the proteins identified using these techniques will help to refine protocols for the production of dopamine neurons in vitro.

scholarly journals Using bioinformatics for the identification of key peptides to engineer dopamine neurons. Towards a therapy for Parkinson’s disease.

2018 ◽  
Author(s):  
William D Mitchell ◽  
Rowan P Orme ◽  
Sarah R Hart ◽  
Rosemary A Fricker
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Absolute Quantification ◽  
Dopamine Neurons ◽  
Neuron Development ◽  
Expression Levels ◽  
Relative Expression ◽  
Online Databases ◽  
Isobaric Tagging

Parkinson’s disease is a widespread condition caused by degeneration of dopamine neurons in the midbrain. A number of proteins are known to be important to signalling mechanisms present in the midbrain during natural dopamine neuron development, and may be utilised to better produce dopamine neurons in vitro. Relative expression levels of proteins were obtained from substantia nigra tissue of rats from embryonic days E11 through E14 using isobaric tagging for relative and absolute quantification. This project analysed the dataset obtained, with an emphasis on relative expression levels of proteins across the four-day period. Bioinformatics searching of online databases reduced the dataset from 3325 proteins to a shortlist of five worthy of further investigation. It is hoped that the proteins identified using these techniques will help to refine protocols for the production of dopamine neurons in vitro.

Angiostatic Proteins and Peptides

2001 ◽  
Vol 11 (4) ◽  
pp. 16 ◽  
Author(s):  
Jessica C. A. Bouma-ter Steege ◽  
Kevin H. Mayo ◽  
Arjan W. Griffioen
Keyword(s):  
Proteins And Peptides
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