A High-Throughput Biological Calorimetry Core

Abstract

Many labs have conventional calorimeters where denaturation and binding experiments are setup and run one at a time. While these systems are highly informative to biopolymer folding and ligand interaction, they require considerable manual intervention for cleaning and setup. As such, the throughput for such setups is limited typically to a few runs a day. With a large number of experimental parameters to explore including different buffers, macromolecule concentrations, temperatures, ligands, mutants, controls, replicates, and instrument tests, the need for high-throughput automated calorimeters is on the rise. Lower sample volume requirements and reduced user intervention time compared to the manual instruments have improved turnover of calorimetry experiments in a high-throughput format where 25 or more runs can be conducted per day. The cost and efforts to maintain high-throughput equipment typically demands that these instruments be housed in a multiuser core facility. We describe here the steps taken to successfully start and run an automated biological calorimetry facility at Pennsylvania State University. Scientists from various departments at Penn State including Chemistry, Biochemistry and Molecular Biology, Bioengineering, Biology, Food Science, and Chemical Engineering are benefiting from this core facility. Samples studied include proteins, nucleic acids, sugars, lipids, synthetic polymers, small molecules, natural products, and virus capsids. This facility has led to higher throughput of data, which has been leveraged into grant support, attracting new faculty hire and has led to some exciting publications.

Membrane Active Peptides and Their Biophysical Characterization

“…In this technique, a monochromatic beam of light encounters a biomolecule solution and fluctuations in the scattered light intensity are analyzed [ 228 , 229 , 230 , 231 ]. Using this approach it is possible to get information on the microsecond timescale about the size distribution of aggregated species in the lipid bilayer [ 231 , 232 ]. For example, the antimicrobial activity of gomesin was related to its ability to induce vesicle aggregation, as detected by DLS [ 146 ].…”

Zn 2+ -Inducible Expression Platform for Synechococcus sp. Strain PCC 7002 Based on the smtA Promoter/Operator and smtB Repressor

“…The YFP concentration was determined using a Direct Detect infrared spectrometer (EMD Millipore, Merck, Darmstadt, Germany). YFP protein concentrations were determined using the amide 1 region of the infrared (IR) spectrum (39,40).…”