Thomas Corso - Academia.edu (original) (raw)
Papers by Thomas Corso
Scientific Reports
Viral vectors represent a bottleneck in the manufacturing of cellular therapies. Electroporation ... more Viral vectors represent a bottleneck in the manufacturing of cellular therapies. Electroporation has emerged as an approach for non-viral transfection of primary cells, but standard cuvette-based approaches suffer from low throughput, difficult optimization, and incompatibility with large-scale cell manufacturing. Here, we present a novel electroporation platform capable of rapid and reproducible electroporation that can efficiently transfect small volumes of cells for research and process optimization and scale to volumes required for applications in cellular therapy. We demonstrate delivery of plasmid DNA and mRNA to primary human T cells with high efficiency and viability, such as > 95% transfection efficiency for mRNA delivery with < 2% loss of cell viability compared to control cells. We present methods for scaling delivery that achieve an experimental throughput of 256 million cells/min. Finally, we demonstrate a therapeutically relevant modification of primary T cells u...
Molecular & Cellular Proteomics, Jun 1, 2013
We report the development and characterization of a novel, vendor-neutral ultra-high pressure-com... more We report the development and characterization of a novel, vendor-neutral ultra-high pressure-compatible (ϳ10,000 p.s.i.) LC-MS source. This device is the first to make automated connections with user-packed capillary traps, columns, and capillary emitters. The source uses plastic rectangular inserts (referred to here as cartridges) where individual components (i.e. trap, column, or emitter) can be exchanged independent of one another in a plug and play manner. Automated robotic connections are made between the three cartridges using linear translation powered by stepper motors to axially compress each cartridge by applying a well controlled constant compression force to each commercial LC fitting. The user has the versatility to tailor the separation (e.g. the length of the column, type of stationary phase, and mode of separation) to the experimental design of interest in a cost-effective manner. The source is described in detail, and several experiments are performed to evaluate the robustness of both the system and the exchange of the individual trap and emitter cartridges. The standard deviation in the retention time of four targeted peptides from a standard digest interlaced with a soluble Caenorhabditis elegans lysate ranged between 3.1 and 5.3 s over 3 days of analyses. Exchange of the emitter cartridge was found to have an insignificant effect on the abundance of various peptides. In addition, the trap cartridge can be replaced with minimal effects on retention time (<20 s).
Analytical Chemistry, Jan 30, 2017
As compared to conventional high performance liquid chromatography (HPLC) techniques, nanoflow HP... more As compared to conventional high performance liquid chromatography (HPLC) techniques, nanoflow HPLC exhibits increased sensitivity and limits of detection. However, nanoflow HPLC suffers from low throughput due to instrument failure (e.g. fitting fatigue and trapping column failure), limiting the utility of the technique for clinical and industrial applications. To increase the robustness of nanoflow HPLC, we have developed and tested a trapping column exchanging robot for autonomous interchange of trapping columns. This robot makes reproducible, automated connections between the active trapping column and the rest of the HPLC system. The intertrapping column retention time is shown to be sufficiently reproducible for scheduled selected reaction monitoring assays to be performed on different trapping columns without rescheduling the selection windows.
![Research paper thumbnail of [3‐13C] γ‐linolenic acid: A new probe for 13C nuclear magnetic resonance studies of arachidonic acid synthesis in the suckling rat](https://attachments.academia-assets.com/110507014/thumbnails/1.jpg)
](Lipids, 1997
Our objective was to develop a suitable probe to study metabolism of polyunsaturated fatty acids ... more Our objective was to develop a suitable probe to study metabolism of polyunsaturated fatty acids by 13C nuclear magnetic resonance (NMR) in the suckling rat pup. [3‐13C] γ‐Linolenic acid was chemically synthesized, and a 20 mg (Experiment 1) or 5 mg (Experiment 2) dose was injected into the stomachs of 6–10‐day‐old suckling rat pups that were then killed over a 192 h (8 d) time course. 13C NMR showed that 13C in γ‐linolenate peaked in liver total lipids by 12‐h post‐dosing and that [5‐13C]‐arachidonic acid peaked in both brain and liver total lipids 48–96 h post‐dosing. 13C enrichment in brain γ‐linolenic acid was not detected by NMR, but gas chromatography‐combustion‐isotope ratio mass spectrometry showed that its mass enrichment in brain phospholipids at 48–96 h post‐dosing was 1–2% of that in brain arachidonic acid. 13C was present in liver and brain cholesterol and in perchloric acid‐extractable water‐soluble metabolites in the brain, liver and carcass. We conclude that low but ...
Methods: • Two standard phosphoproteins, bovine α -casein and β-casein, were used to demonstrate ... more Methods: • Two standard phosphoproteins, bovine α -casein and β-casein, were used to demonstrate the method. • Calf intestinal phosphatase (CIP) was used to treat the α -casein digest. • The NanoMate and ESI Chip™ were used to infuse the samples into the LTQ for DDA analysis followed by automated neutral-loss scanning to identify phosphorylation sites. Results: • Definitive identification of phosphorylation sites of both α -casein and β-casein digests using chip-based nanoESI/MS n with automated neutralloss scanning was demonstrated. • Phosphatase treatment prior to MS analysis is required for phosphorylation mapping for multiple phosphate groups in a single peptide.
Lab Chip, 2014
Fabrication and operation of a microtensiometer to sense environmental water status and measure e... more Fabrication and operation of a microtensiometer to sense environmental water status and measure equation of state data under tension.
Absorption and metabolism of [13C]9-cis-f3-carotene ([13C]9cf3C) was studied in three subjects af... more Absorption and metabolism of [13C]9-cis-f3-carotene ([13C]9cf3C) was studied in three subjects after a single oral dose. Subjects given 1.0 mg e3C]f3-carotene (mean: 99.4% 9-cisf3-carotene, 0.6% all~trans-f3-carotene; dose A) had substantial concentrations of [13C]all-trans-f3-carotene ([13C]trf3C) and [13C]all-trans retinol ([13C]retinol) but very low concentrations of [13C]cis-f3-carotene ([13C]cisf3c) in saponified plasma 5 h after dosing, as determined by HPLC and isotope-ratio mass spectrometry. There was no evidence of appreciable absorption of [13C]9cis retinol. To determine the proportion of [13C]trf3C and [13C]retinol derived from [13C]9cf3C, a second set of studies in the same subjects was performed with the same isomeric composition except with 13C labeling only in all-trans-f3-carotene (dose B). The results indicated that> 95% of plasma [13C]trf3C and [13C]retinol observed after dose A was derived from [13C]9cf3C. The concentrations of [13C]trf3C observed, in excess o...
L'invention concerne un dispositif d'electronebulisation (100), un dispositif de chromato... more L'invention concerne un dispositif d'electronebulisation (100), un dispositif de chromatographie en phase liquide et un dispositif d'electronebulisation et de chromatographie en phase liquide. Le dispositif d'electronebulisation (100) comporte un substrat (102) qui definit un canal (104) entre un orifice d'entree (106) sur une surface d'injection (108) et un orifice de sortie sur une surface d'ejection (112), une buse (110) definie par une partie en retrait (114) par rapport a la surface d'ejection (112) entourant l'orifice de sortie, et une electrode pour l'application d'un potentiel electrique au substrat (102) afin d'optimiser et de generer une electronebulisation. L'orifice de sortie du dispositif de chromatographie en phase liquide peut etre couple de facon homogene a l'orifice d'entree (106) du dispositif d'electronebulisation (100) pour constituer un seul et unique systeme integre.
Analytical Chemistry
As compared to conventional high performance liquid chromatography (HPLC) techniques, nanoflow HP... more As compared to conventional high performance liquid chromatography (HPLC) techniques, nanoflow HPLC exhibits increased sensitivity and limits of detection. However, nanoflow HPLC suffers from low throughput due to instrument failure (e.g. fitting fatigue and trapping column failure), limiting the utility of the technique for clinical and industrial applications. To increase the robustness of nanoflow HPLC, we have developed and tested a trapping column exchanging robot for autonomous interchange of trapping columns. This robot makes reproducible, automated connections between the active trapping column and the rest of the HPLC system. The intertrapping column retention time is shown to be sufficiently reproducible for scheduled selected reaction monitoring assays to be performed on different trapping columns without rescheduling the selection windows.
Scientific Reports
Viral vectors represent a bottleneck in the manufacturing of cellular therapies. Electroporation ... more Viral vectors represent a bottleneck in the manufacturing of cellular therapies. Electroporation has emerged as an approach for non-viral transfection of primary cells, but standard cuvette-based approaches suffer from low throughput, difficult optimization, and incompatibility with large-scale cell manufacturing. Here, we present a novel electroporation platform capable of rapid and reproducible electroporation that can efficiently transfect small volumes of cells for research and process optimization and scale to volumes required for applications in cellular therapy. We demonstrate delivery of plasmid DNA and mRNA to primary human T cells with high efficiency and viability, such as > 95% transfection efficiency for mRNA delivery with < 2% loss of cell viability compared to control cells. We present methods for scaling delivery that achieve an experimental throughput of 256 million cells/min. Finally, we demonstrate a therapeutically relevant modification of primary T cells u...
Molecular & Cellular Proteomics, Jun 1, 2013
We report the development and characterization of a novel, vendor-neutral ultra-high pressure-com... more We report the development and characterization of a novel, vendor-neutral ultra-high pressure-compatible (ϳ10,000 p.s.i.) LC-MS source. This device is the first to make automated connections with user-packed capillary traps, columns, and capillary emitters. The source uses plastic rectangular inserts (referred to here as cartridges) where individual components (i.e. trap, column, or emitter) can be exchanged independent of one another in a plug and play manner. Automated robotic connections are made between the three cartridges using linear translation powered by stepper motors to axially compress each cartridge by applying a well controlled constant compression force to each commercial LC fitting. The user has the versatility to tailor the separation (e.g. the length of the column, type of stationary phase, and mode of separation) to the experimental design of interest in a cost-effective manner. The source is described in detail, and several experiments are performed to evaluate the robustness of both the system and the exchange of the individual trap and emitter cartridges. The standard deviation in the retention time of four targeted peptides from a standard digest interlaced with a soluble Caenorhabditis elegans lysate ranged between 3.1 and 5.3 s over 3 days of analyses. Exchange of the emitter cartridge was found to have an insignificant effect on the abundance of various peptides. In addition, the trap cartridge can be replaced with minimal effects on retention time (<20 s).
Analytical Chemistry, Jan 30, 2017
As compared to conventional high performance liquid chromatography (HPLC) techniques, nanoflow HP... more As compared to conventional high performance liquid chromatography (HPLC) techniques, nanoflow HPLC exhibits increased sensitivity and limits of detection. However, nanoflow HPLC suffers from low throughput due to instrument failure (e.g. fitting fatigue and trapping column failure), limiting the utility of the technique for clinical and industrial applications. To increase the robustness of nanoflow HPLC, we have developed and tested a trapping column exchanging robot for autonomous interchange of trapping columns. This robot makes reproducible, automated connections between the active trapping column and the rest of the HPLC system. The intertrapping column retention time is shown to be sufficiently reproducible for scheduled selected reaction monitoring assays to be performed on different trapping columns without rescheduling the selection windows.
Lipids, 1997
Our objective was to develop a suitable probe to study metabolism of polyunsaturated fatty acids ... more Our objective was to develop a suitable probe to study metabolism of polyunsaturated fatty acids by 13C nuclear magnetic resonance (NMR) in the suckling rat pup. [3‐13C] γ‐Linolenic acid was chemically synthesized, and a 20 mg (Experiment 1) or 5 mg (Experiment 2) dose was injected into the stomachs of 6–10‐day‐old suckling rat pups that were then killed over a 192 h (8 d) time course. 13C NMR showed that 13C in γ‐linolenate peaked in liver total lipids by 12‐h post‐dosing and that [5‐13C]‐arachidonic acid peaked in both brain and liver total lipids 48–96 h post‐dosing. 13C enrichment in brain γ‐linolenic acid was not detected by NMR, but gas chromatography‐combustion‐isotope ratio mass spectrometry showed that its mass enrichment in brain phospholipids at 48–96 h post‐dosing was 1–2% of that in brain arachidonic acid. 13C was present in liver and brain cholesterol and in perchloric acid‐extractable water‐soluble metabolites in the brain, liver and carcass. We conclude that low but ...
Methods: • Two standard phosphoproteins, bovine α -casein and β-casein, were used to demonstrate ... more Methods: • Two standard phosphoproteins, bovine α -casein and β-casein, were used to demonstrate the method. • Calf intestinal phosphatase (CIP) was used to treat the α -casein digest. • The NanoMate and ESI Chip™ were used to infuse the samples into the LTQ for DDA analysis followed by automated neutral-loss scanning to identify phosphorylation sites. Results: • Definitive identification of phosphorylation sites of both α -casein and β-casein digests using chip-based nanoESI/MS n with automated neutralloss scanning was demonstrated. • Phosphatase treatment prior to MS analysis is required for phosphorylation mapping for multiple phosphate groups in a single peptide.
Lab Chip, 2014
Fabrication and operation of a microtensiometer to sense environmental water status and measure e... more Fabrication and operation of a microtensiometer to sense environmental water status and measure equation of state data under tension.
Absorption and metabolism of [13C]9-cis-f3-carotene ([13C]9cf3C) was studied in three subjects af... more Absorption and metabolism of [13C]9-cis-f3-carotene ([13C]9cf3C) was studied in three subjects after a single oral dose. Subjects given 1.0 mg e3C]f3-carotene (mean: 99.4% 9-cisf3-carotene, 0.6% all~trans-f3-carotene; dose A) had substantial concentrations of [13C]all-trans-f3-carotene ([13C]trf3C) and [13C]all-trans retinol ([13C]retinol) but very low concentrations of [13C]cis-f3-carotene ([13C]cisf3c) in saponified plasma 5 h after dosing, as determined by HPLC and isotope-ratio mass spectrometry. There was no evidence of appreciable absorption of [13C]9cis retinol. To determine the proportion of [13C]trf3C and [13C]retinol derived from [13C]9cf3C, a second set of studies in the same subjects was performed with the same isomeric composition except with 13C labeling only in all-trans-f3-carotene (dose B). The results indicated that> 95% of plasma [13C]trf3C and [13C]retinol observed after dose A was derived from [13C]9cf3C. The concentrations of [13C]trf3C observed, in excess o...
L'invention concerne un dispositif d'electronebulisation (100), un dispositif de chromato... more L'invention concerne un dispositif d'electronebulisation (100), un dispositif de chromatographie en phase liquide et un dispositif d'electronebulisation et de chromatographie en phase liquide. Le dispositif d'electronebulisation (100) comporte un substrat (102) qui definit un canal (104) entre un orifice d'entree (106) sur une surface d'injection (108) et un orifice de sortie sur une surface d'ejection (112), une buse (110) definie par une partie en retrait (114) par rapport a la surface d'ejection (112) entourant l'orifice de sortie, et une electrode pour l'application d'un potentiel electrique au substrat (102) afin d'optimiser et de generer une electronebulisation. L'orifice de sortie du dispositif de chromatographie en phase liquide peut etre couple de facon homogene a l'orifice d'entree (106) du dispositif d'electronebulisation (100) pour constituer un seul et unique systeme integre.
Analytical Chemistry
As compared to conventional high performance liquid chromatography (HPLC) techniques, nanoflow HP... more As compared to conventional high performance liquid chromatography (HPLC) techniques, nanoflow HPLC exhibits increased sensitivity and limits of detection. However, nanoflow HPLC suffers from low throughput due to instrument failure (e.g. fitting fatigue and trapping column failure), limiting the utility of the technique for clinical and industrial applications. To increase the robustness of nanoflow HPLC, we have developed and tested a trapping column exchanging robot for autonomous interchange of trapping columns. This robot makes reproducible, automated connections between the active trapping column and the rest of the HPLC system. The intertrapping column retention time is shown to be sufficiently reproducible for scheduled selected reaction monitoring assays to be performed on different trapping columns without rescheduling the selection windows.