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.
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
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