Protein Refolding and Disaggregation Display: ASC DESC Sort By Title Date Ordering Confronting high-throughput protein refolding using high pressure and solution screens M. Walid Qoronfleha,*, Lyndal K. Hesterbergb, Matthew B. Seefeldtb aUniversity of Michigan and Core Technology Alliance-CTA, 1024 Wolverine Tower, 3003 State Street, Ann Arbor, MI 48109-1274, USA bBaroFold Inc., 1825 33rd Street, Boulder, CO 80301, USA Download Details High hydrostatic pressure as a tool to study protein aggregation and amyloidosis. Randolph TW, Seefeldt M, Carpenter JF. Department of Chemical Engineering, University of Colorado, Boulder, CO 80309, USA. Download Details High-pressure refolding of bikunin: efficacy and thermodynamics. Seefeldt MB1, Ouyang J, Froland WA, Carpenter JF, Randolph TW. Department of Chemical and Biological Engineering, Center for Pharmaceutical Biotechnology, ECCH 111, University of Colorado, Boulder, CO 80309-0424, USA. Download Details High-pressure studies of aggregation of recombinant human interleukin-1 receptor antagonist: thermodynamics, kinetics, and application to accelerated formulation studies. Seefeldt MB1, Kim YS, Tolley KP, Seely J, Carpenter JF, Randolph TW Department of Chemical and Biological Engineering, Center for Pharmaceutical Biotechnology, ECCH 111, University of Colorado, Boulder, CO 80309-0242, USA. Download Details Specific volume and adiabatic compressibility measurements of native and aggregated recombinant human interleukin-1 receptor antagonist: density differences enable pressure-modulated refolding. Seefeldt MB1, Crouch C, Kendrick B, Randolph TW. Department of Chemical and Biological Engineering, University of Colorado, Boulder, Colorado 80309, USA. Download Details Application of high hydrostatic pressure to dissociate aggregates and refold proteins. Seefeldt MB1, Rosendahl MS, Cleland JL, Hesterberg LK. BaroFold Inc., Boulder, CO 80301, USA. Download Details Disaggregation of Proinsulin Inclusion Bodies by High Pressure FEATURED Introduction Protein expression in E. coli is an efficient and commonly used method to generate large quantities of protein for research or therapeutic applications. Unfortunately, proteins expressed at high levels in E. coli are often packaged into inclusion bodies (IBs). These tightly-packed structures have the advantage of being composed of almost pure expressed protein, but the serious disadvantage that the protein is so tightly aggregated that high concentrations of chaotropes or detergents are required to extract soluble protein from the aggregates. These solubilization reagents must then be diluted or removed by buffer exchange, so that the extracted protein can be refolded into its native, functional conformation. High hydrostatic pressure has shown promise as a means of disaggregating and solubilizing protein aggregates using relatively mild buffer conditions [1-4]. By disaggregating IBs without the high levels of denaturants required under conventional conditions, subsequent protein refolding can be improved. Here we report that high hydrostatic pressure can be used to efficiently disaggregate proinsulin inclusion bodies in order to extract soluble proinsulin protein. This disaggregation can be carried out in mild buffer conditions at ambient temperature in as little as 5 minutes at 45kpsi. Download View Details Rapid Refolding of Protein from Inclusion Bodies by High Pressure FEATURED Introduction Protein expression in E. coli is an efficient and commonly used method to generate large quantities of protein forresearch or therapeutic applications. Unfortunately, proteins expressed at high levels in E. coli are oftenpackaged into inclusion bodies (IBs). These tightly packed structures have the advantage of being composed ofalmost pure expressed protein, but the serious disadvantage that the protein is so tightly aggregated that highconcentrations of chaotropes or detergents are required to extract soluble protein from the aggregates. Thesesolubilization reagents must then be diluted or removed by buffer exchange, so that the extracted protein can berefolded into its native, functional conformation. Download View Details