Open Access. Powered by Scholars. Published by Universities.®
Articles 1 - 3 of 3
Full-Text Articles in Medicine and Health Sciences
Identification Of Highly Expressed, Soluble Proteins Using An Improved, High-Throughput Pooled Orf Expression Technology, Timothy Waybright, William Gillette, Dominic Esposito, Robert Stephens, David Lucas, James Hartley, Timothy D. Veenstra
Identification Of Highly Expressed, Soluble Proteins Using An Improved, High-Throughput Pooled Orf Expression Technology, Timothy Waybright, William Gillette, Dominic Esposito, Robert Stephens, David Lucas, James Hartley, Timothy D. Veenstra
Pharmaceutical Sciences Faculty Publications
This article describes an improved pooled open reading frame (ORF) expression technology (POET) that uses recombinational cloning and solution-based tandem mass spectrometry (MS/MS) to identify ORFs that yield high levels of soluble, purified protein when expressed in Escherichia coli. Using this method, three identical pools of 512 human ORFs were subcloned, purified, and transfected into three separate E. coli cultures. After bulk expression and purification, the proteins from the three separate pools were digested into tryptic peptides. Each of these samples was subsequently analyzed in triplicate using reversed-phase high-performance liquid chromatography (LC) coupled directly online with MS/MS. The abundance of …
Two-Dimensional Polyacrylamide Gel Electrophoresis (2d-Page): Advances And Perspectives, Haleem Issaq, Timothy D. Veenstra
Two-Dimensional Polyacrylamide Gel Electrophoresis (2d-Page): Advances And Perspectives, Haleem Issaq, Timothy D. Veenstra
Pharmaceutical Sciences Faculty Publications
The recent trend in science is to assay as many biological molecules as possible within a single experiment. This trend is evident in proteomics where the aim is to characterize thousands of proteins within cells, tissues, and organisms. While advances in mass spectrometry have been critical, developments made in two-dimensional PAGE (2D-PAGE) have also played a major role in enabling proteomics. In this review, we discuss and highlight the advances made in 2D-PAGE over the past 25 years that have made it a foundational tool in proteomic research.
Mass Spectrometry: M/Z 1983-2008, Ming Zhou, Timothy D. Veenstra
Mass Spectrometry: M/Z 1983-2008, Ming Zhou, Timothy D. Veenstra
Pharmaceutical Sciences Faculty Publications
While definitely not a new technology, mass spectrometry (MS) has seen incredible growth over the past 25 years. Mass spectrometry has rapidly evolved to the forefront of analytical techniques; its ability to analyze proteins is the major driving force in the field of proteomics. MS instrumentation has increased approximately 5-fold in sensitivity every three years. The level of performance that is achievable with MS today allows scientists to study proteins in ways that were inconceivable a quarter century ago. This review of the history of MS over the past 25 years is timely in that it encompasses two of the …