Robustness of nanofiltration for increasing the viral safety margin of biological products.
In this study, the virus-removal capacity of nanofiltration was assessed using validated laboratory scale models on a wide range of viruses (pseudorabies virus; human immunodeficiency virus; bovine viral diarrhea virus; West Nile virus; hepatitis A virus; murine encephalomyocarditis virus; and porcine parvovirus) with sizes from 18 nm to 200 nm and applying the different process conditions existing in a number of Grifols' plasma-derived manufacturing processes (thrombin, α1-proteinase inhibitor, Factor IX, antithrombin, plasmin, intravenous immunoglobulin, and fibrinogen). Spiking experiments (n = 133) were performed in process intermediate products, and removal was subsequently determined by infectivity titration. Reduction Factor (RF) was calculated by comparing the virus load before and after nanofiltration under each product purification condition. In all experiments, the RFs were close to or greater than 4 log10 (>99.99% of virus elimination). RF values were not significantly affected by the process conditions within the limits assayed (pH, ionic strength, temperature, filtration ratio, and protein concentration). The virus-removal capacity of nanofiltration correlated only with the size of the removed agent. In conclusion, nanofiltration, as used in the manufacturing of several Grifols' products, is consistent, robust, and not significantly affected by process conditions.
Research and Development Area, Biologics Industrial Group, Grifols, Parets del Vallès, Barcelona, Spain. Electronic address: santiago.caballero@grifols.com.
Santiago
S
Caballero
Research and Development Area, Biologics Industrial Group, Grifols, Parets del Vallès, Barcelona, Spain.
José M
JM
Diez
Research and Development Area, Biologics Industrial Group, Grifols, Parets del Vallès, Barcelona, Spain.
Francisco J
FJ
Belda
Research and Development Area, Biologics Industrial Group, Grifols, Parets del Vallès, Barcelona, Spain.
Magdalena
M
Otegui
Research and Development Area, Biologics Industrial Group, Grifols, Los Angeles, CA, USA.
Steven
S
Herring
Research and Development Area, Biologics Industrial Group, Grifols, Research Triangle Park, NC, USA.
Nathan J
NJ
Roth
Research and Development Area, Biologics Industrial Group, Grifols, Research Triangle Park, NC, USA.
Douglas
D
Lee
Research and Development Area, Biologics Industrial Group, Grifols, Parets del Vallès, Barcelona, Spain.
Rodrigo
R
Gajardo
Research and Development Area, Biologics Industrial Group, Grifols, Parets del Vallès, Barcelona, Spain.
Juan I
JI
Jorquera
D001688:Biological Products / Q000009:adverse effects*; D012449:Safety; D014462:Ultrafiltration / Q000379:methods / Q000592:standards*; D014780:Viruses / Q000302:isolation & purification*
D001688:Biological Products
Nanofiltration; Plasma-derived product; Viral safety