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Enhancing the possibilities of LC-MS/MS for the absolute quantification of proteins in biological samples

27 October 2015

The research of Klaas Bronsema demonstrates that the performance of LC-MS/MS can be in accordance with international guidelines, and that these method is a serious alternative for ligand binding essays for the quantification of proteins in complex biological samples down to the low pg/mL level.

Protein pharmaceuticals or so called “biopharmaceuticals” form a class of innovative therapeutic agents that are rapidly increasing in relevance for health care. In order to develop these large molecules into effective and safe drugs, there is a growing demand for the sensitive and reliable determination of the concentrations of these proteins in  biological fluids such as plasma and serum to support (pre)-clinical research. In this field, ligand binding assays, are currently the standard analytical technique, but in recent years, there is a trend towards the use of liquid chromatography hyphenated with (tandem) mass spectrometry (LC-MS/MS).

One of the main advantages of using LC-MS/MS for quantification of proteins is the possibility of using internal standards, which increases the accuracy and precision of the method. The thesis includes a review of the literature in which eight different approaches for internal standardization are identified and described. In the next chapter, the ability of these approaches to correct for experimental variability is compared by using them in an LC-MS/MS method for   the biopharmaceutical protein salmon calcitonin. Next, the development, validation and application of analytical methods based on LC-MS/MS for biopharmaceutical  and endogenous proteins (biomarkers) in plasma is described.


PhD ceremony:

Mr K.J. Bronsema


October 30, 2015




prof. dr. N.C. van de Merbel , prof. dr. R.P.H. (Rainer) Bischoff


Academy building RUG


Mathematics and Natural Sciences

Last modified:05 April 2017 07.21 a.m.
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