Recombinant protein overexpression of large proteins in bacteria
often results in insoluble and misfolded proteins directed to
inclusion bodies. We report the application of shear stress in
micrometer-wide, thin fluid films to refold boiled hen egg
white lysozyme, recombinant hen egg white lysozyme, and
recombinant caveolin-1. Furthermore, the approach allowed
refolding of a much larger protein, cAMP-dependent protein
kinase A (PKA). The reported methods require only minutes,
which is more than 100 times faster than conventional overnight
dialysis. This rapid refolding technique could significantly
shorten times, lower costs, and reduce waste streams associated
with protein expression for a wide range of industrial and
research applications.