title: Protein denaturation by combined effect of shear and air-liquid interface.
abstract: The effect of shear alone on the aggregation of recombinant human growth hormone (rhGH) and recombinant human deoxyribonuclease (rhDNase) has been found to be insignificant. This study focused on the synergetic effect of shear and gas-liquid interface on these two model proteins. Two shearing systems, the concentric-cylinder shear device (CCSD) and the rotor/stator homogenizer, were used to generate high shear (> 10(6)) in aqueous solutions in the presence of air. High shear in the presence of an air-liquid interface had no major effect on rhDNase but caused rhGH to form noncovalent aggregates. rhGH aggregation was induced by the air-liquid interface and was found to increase with increasing protein concentration and the air-liquid interfacial area. The aggregation was irreversible and exhibited a first-order kinetics with respect to the protein concentration and air-liquid interfacial area. Shear and shear rate enhanced the interaction because of its continuous generation of new air-liquid interfaces. In the presence of a surfactant, aggregation could be delayed or prevented depending upon the type and the concentration of the surfactant. The effect of air-liquid interface on proteins at low shear was examined using a nitrogen bubbling method. We found that foaming is very detrimental to rhGH even though the shear involved is low. The use of anti-foaming materials could prevent rhGH aggregation during bubbling. The superior stability exhibited by rhDNase may be linked to the higher surface tension and lower foaming tendency of its aqueous solution. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 54: 503-512, 1997.
authors.affiliation: Department of Pharmaceutical Research and Development, Genentech, Inc. 460 Point San Bruno Boulevard, South San Francisco, CA 94080, USA. maa.yuh-fun@gene.com <maa.yuh-fun@gene.com>
authors.forename: Y F
authors.initials: YF
authors.lastname: Maa
authors.forename: C C
authors.initials: CC
authors.lastname: Hsu