IBT Protein Production, Characterization, and Molecular Interaction Core-Houston, TX
Protein Production, Characterization and Molecular Interaction (PPCMI) at Texas A&M Institute of Biosciences & Technology is dedicated to providing expression and purification services of functional proteins at a scale that meets the quantity and purity benchmarks for structural, biophysical, biochemical, and therapeutics studies. Services range from the scale-up of existing protocols to the development of de novo expression and purification strategies. Currently, the PPC employs bacteria and yeast host systems with a view to expanding into mammalian host systems. The PPC specializes in systematic method development for expressing and purifying challenging proteins. Currently, the core facility is equipped with an ÄKTA Pure FPLC system, a Microfluidics M-110P cell homogenizer, and a cell culture suite. In addition, the PPC is furnished with all the standard laboratory equipment like refrigerated console shaker/incubators, an Avanti J-E centrifuge, Optima XPN-80 ultracentrifuge, and an Eppendorf D30 UV-vis spectrophotometer to expedite scale-up of protein production and purification. Also, the PPC is equipped with a Mosquito LCP and a Dragonfly screen optimizer for automated protein crystallization screening, optimization, and scale-up, as well as a Biacore T200 (SPR) and a MicroCal PEAQ-ITC for high-quality characterization of molecular interactions in terms of kinetics, affinity, specificity, comparability, immunogenicity, and thermodynamics.
Equipment:
- New Brunswick’s I26 incubated and refrigerated console shakers: capable of large scale expression of recombinant proteins in bacterial and yeast expression systems.
- Microfluidics’ Microfluidizer M 110P: capable of large scale and high-efficiency cell lysis.
- Beckman Coulter’s Avanti J-E centrifuge and Optima XPN-80 ultracentrifuge: capable of large scale and high-efficiency cell lysate separation and membrane preparation.
- GE Healthcare’s ÄKTA Pure chromatography system: a flexible and intuitive purification system for proteins, peptides, and nucleic acids from microgram levels to tens of grams of target product in research applications.
- TTP Labtech’s Mosquito LCP: automated protein crystallization screening, optimization, and scale-up.
- TTP Labtech’s Dragonfly screen optimizer: enables complex assay gradients or optimization screens to be rapidly and accurately prepared, even with viscous liquids, while eliminating cross-contamination.
- GE Healthcare’s Biacore T200: a versatile system for high-quality characterization of molecular interactions in terms of kinetics, affinity, specificity, comparability, immunogenicity, and thermodynamics.
- Marlvern’s MicroCal PEAQ-ITC: allows direct, label-free in solution measurement of binding affinity and thermodynamics in a single experiment, enabling the accurate determination of binding constants, reaction stoichiometry, enthalpy, and entropy; provides a complete thermodynamic profile of the molecular interaction.
The list below summarizes our comprehensive service offerings at the core facility:
- Custom cloning service (Genomic DNA clone; cDNA clone; genetic engineering)
- Site-directed mutagenesis (Single mutation and multiple mutations in a single construct; library of random mutants)
- Expression optimization tests
- Pilot scale protein expression and purification
- Large-scale protein production
- Co-expression and purification of protein complexes
- Protein refolding from inclusion bodies
- Tag removal for purified recombinant proteins
- SDS-PAGE & Western Blot assays for expressed recombinant proteins
- Size-exclusion chromatography (GE Healthcare’s prepacked SEC columns from analytic scale Superdex increase to production grade Superdex) on ÄKTA Pure system
- Affinity chromatography (GE Healthcare’s prepacked affinity columns, i.e. HisTrap HP, StrepTrap HP, GSTrap HP, MBPTrap HP) on ÄKTA Pure system
- Endotoxin removal for purified recombinant proteins
- Multiple-step protein purification on ÄKTA Pure system
- Protein activity assay
- Protein crystallization screening, optimization, and scale-up; customers also can rent time on Mosquito LCP and Dragonfly screen optimizer with our supervision and assistance.
- Spectroscopy based biophysics assays (CD, ITC and SPR); customers also can rent time on MicroCal PEAQ-ITC and Biacore T200 with our supervision and assistance.
