The Development and Implementation of Single Molecule Protein Assays for Applications in Early Cancer Diagnostics and Single Cell Studies.
Abstract: Earlier detection of diseases, such as cancer, often leads to
improved prognosis. Therefore, the ability to efficiently, accurately, sensitively, and
non-invasively screen for cancer at the earliest stages possible is of paramount
importance. Furthermore, a better understanding of the basic biology of molecular
mechanisms, cellular pathways, and cellular heterogeneity may lead to mor... read moree personalized and
efficient therapies. In order to achieve this task and to also detect rare cells that may
also enable earlier disease detection, specific biomarkers must be studied at single
molecule resolution within individual cells. This thesis focuses on the detection of
protein biomarkers at ultralow levels in serum for early cancer detection and for
fundamental single cell studies using single molecule protein counting technology. This
thesis describes the technology, single molecule arrays (SiMoA), that is utilized
throughout the included works and discusses the fundamental kinetics behind the method.
Chapter 3 describes how a biomarker can be detected at ultralow concentrations in serum
prior to palpable tumor formation using a mouse model, indicating the utility of SiMoA as
an early cancer detection tool. Chapter 4 describes the use of SiMoA in a panel of breast
cancer protein biomarkers and examines the utility of these biomarkers in detecting early
stage breast cancer from serum samples by using supervised multivariate regression models.
Chapter 5 demonstrates the use of SiMoA technology as a straightforward approach for
counting single protein molecules within single cells. Chapter 6 includes work towards
creating a breast cancer mouse model for studying early cancer progression. One appendix is
included that includes detailed patient information relevant to Chapter 4.
Thesis (Ph.D.)--Tufts University, 2015.
Submitted to the Dept. of Chemistry.
Advisor: David Walt.
Committee: Samuel Thomas, Joshua Kritzer, and Marsha Moses.
Keywords: Chemistry, and Analytical chemistry.read less