Bioanalysis of therapeutic macromolecules in biological matrices has been a challenging task due to the complexity of both sample matrices and characteristics of drug molecules. Their high potency normally results in low levels in biological samples in PK/PD studies. Therefore a sensitive bioanalytical method is required to monitor low ng/mL or even pg/mL levels in blood plasma for drug development. With recent advances in instrument technology and bioanalytical science, LC-MS has gradually become the preferred analytical technique for many macromolecular analyses, since it allows rapid, sensitive and selective measurements not only for small molecules, but also for moderately large molecules such as polypeptides and oligonucleotides.
In this research work, selective and sensitive LC-MS -based approaches for polypeptides and oligonucleotides in biological matrix have been investigated and discussed in chapter 4 and 5. In order to minimize matrix effect, eliminate endogenous interferences and enhance selectivity as well as sensitivity, the current bioanalysis strategies involve three major steps of the enhancement in LC-MS/MS: sample preparation and enrichment, liquid chromatography and mass spectrometry. Many unique and practical strategies have been proposed and examined, to solve critical issues as ionization efficiency, nonspecific binding, and matrix interference. These approaches have been clearly demonstrated through examples: analysis of glucagon and its analogs (MW, 3000-4000 Da), as well as oligonucleotide drugs (MW, 6000-8000 Da), that LC-MS/MS-based methods have great advantages, and could play more and more important roles in macromolecule analysis in the future.
|Commitee:||Gupta, Pardeep, McEwen, Charles, Schaefer, Fred|
|School:||University of the Sciences in Philadelphia|
|Department:||Chemistry and Biochemistry|
|School Location:||United States -- Pennsylvania|
|Source:||DAI-B 76/03(E), Dissertation Abstracts International|
|Keywords:||Bioanalysis, Glucagon, Matrix effect, Multiple reactions, Oligonucleotide, Polypeptide, Therapeutic macromolecules|
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