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|Title:||Algorithms for the Automated Absolute Quantification of Diagnostic Markers in Complex Proteomics Samples|
|Authors:||GRÖPL Clemens; LANGE Eva; REINERT Knut; KOHLBACHER Oliver; STURM Marc; HUBER Christian; MAYR Bettina; KLEIN CHRISTOPH|
|Citation:||Lecture Notes in Bioinformatics vol. 3695 p. 151-162|
|Type:||Articles in periodicals and books|
|Abstract:||HPLC-ESI-MS is rapidly becoming an established standard method for shotgun proteomics. Currently, its major drawbacks are two-fold: quantification is mostly limited to relative quantification and the large amount of data produced by every individual experiment can make manual analysis quite difficult. Here we present a new, combined experimental and algorithmic approach to absolutely quantify proteins from samples with unprecedented precision. We apply the method to the analysis of myoglobin in human blood serum, which is an important diagnostic marker for myocardial infarction. Our approach was able to determine the absolute amount of myoglobin in a serum sample through a series of standard addition experiments with a relative error of 2.5%. Compared to a manual analysis of the same dataset we could improve the precision and conduct it in a fraction of the time needed for the manual analysis. We anticipate that our automatic quantitation method will facilitate further absolute or relative quantitation of even more complex peptide samples. The algorithm was developed using our publically available software framework OpenMS (www.openms.de)|
|JRC Directorate:||Institute for Health and Consumer Protection Historical Collection|
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