Presentation Type
Poster
Quantifying protein expression associated with drug resistance in single myeloma cells
Abstract
Multiple myeloma is an incurable blood cancer; it is known to develop resistance to chemotherapy. Quantification and identification of proteins in single cells can relate protein expression to the development of drug resistance as well as assess heterogeneity between individual cells in both naïve and drug-resistant cell lines. A pipeline method using liquid chromatography coupled to mass spectrometry for discovery and quantification was developed to measure protein expression in individual cells from multiple myeloma and melphalan-resistant multiple myeloma cell lines. Instrument sensitivity was tested with serial dilutions of both cell lines to confirm the possibility of quantifying proteins in individual cells. Serial dilution method and single cell suspension showed similar signal detection rates. A comparison between both cell lines distinguished variability in protein expression associated with drug resistance. A comparison was conducted between the serial dilution and two methods that use capillary action or flow cytometry to isolate a single cell. Single cell protein quantification is possible with mass spectrometry; with additional development, it may be translated to patient assessment. These efforts may provide the first steps for a new generation of extremely sensitive tools for biomarker monitoring that can be deployed in finding the molecular basis for personalized medicine.
Categories
Biomedical Sciences
Research Type
Thesis
Mentor Information
Dr. John Koomen
Quantifying protein expression associated with drug resistance in single myeloma cells
Multiple myeloma is an incurable blood cancer; it is known to develop resistance to chemotherapy. Quantification and identification of proteins in single cells can relate protein expression to the development of drug resistance as well as assess heterogeneity between individual cells in both naïve and drug-resistant cell lines. A pipeline method using liquid chromatography coupled to mass spectrometry for discovery and quantification was developed to measure protein expression in individual cells from multiple myeloma and melphalan-resistant multiple myeloma cell lines. Instrument sensitivity was tested with serial dilutions of both cell lines to confirm the possibility of quantifying proteins in individual cells. Serial dilution method and single cell suspension showed similar signal detection rates. A comparison between both cell lines distinguished variability in protein expression associated with drug resistance. A comparison was conducted between the serial dilution and two methods that use capillary action or flow cytometry to isolate a single cell. Single cell protein quantification is possible with mass spectrometry; with additional development, it may be translated to patient assessment. These efforts may provide the first steps for a new generation of extremely sensitive tools for biomarker monitoring that can be deployed in finding the molecular basis for personalized medicine.
