Investigation on Fouling Mechanisms for Recombinant Human Growth Hormone Sterile Filtration

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Abstract
During sterile filtration of recombinant human growth hormone solutions, severe membrane fouling was experienced compared to other protein preparations of significantly higher molecular weights and concentrations. This phenomenon was attributed to rhGH aggregation/adsorption occurring in the filter pore. To better understand this phenomenon, we examined several possible fouling mechanisms: (1) pore constriction, (2) adsorption due to nonspecific binding between protein and the membrane, (3) shear-induced adsorption, (4) hydrophobic interface-induced aggregation/adsorption. The protein solutions were sterily filtered using 0.22 mm filters, and their filtration fluxes were monitored. Filtration on the capillary and the noncapillary filters suggested that constraints by pore constriction and tortuosity played only a limited role. Filtration using filters with different degrees of protein binding tendency suggested that nonspecific adsorption was insignificant. The shear stress acting on the protein during filtration was small. RhGH which was intentionally sheared in a high-speed concentrically rotating device did not aggravate fouling tendency, suggesting that the shear-induced adsorption might not be the major fouling mechanism. The dynamic light scattering data showed a trace amount of rhGH aggregates always present in equilibrium with the hydrophobic (air-water and membrane-water) interface. These aggregates tended to be adsorbed to the membrane, and more aggregates were generated presumably due to the equilibrium between aggregates and protein monomers. This adsorption/aggregation process eventually fouled the membrane. When the hydrophobic interface was occupied by surfactant molecules, the equilibration kinetics ceased to generate aggregates, thereby minimizing membrane fouling. This study clarified the cause of such an unusual fouling phenomenon upon microfiltration.