Removal of small nonenveloped viruses by antibody-enhanced nanofiltration during the manufacture of plasma derivatives.
BACKGROUND: Filters with nominal pore sizes in the nanometer range are well-established tools for enhancing the virus safety margins of plasma-derived products, yet intrinsically less successful for smaller viruses such as hepatitis A virus (HAV) and human parvovirus B19 (B19V). The formation of virus-antibody complexes increases the effective size of these smaller viruses and would thus improve their removal by nanofiltration. While the principle of virus removal by antibody-dependent nanofiltration has been demonstrated with animal antisera and viruses spiked into human plasma product intermediates, the significance of these results remains unclear due to the potential contributions of xenoanti-bodies and/or heteroagglutination in such heterologous systems. STUDY DESIGN AND METHODS: The current study investigated antibody-dependent virus removal by nanofiltration in a heterologous animal parvovirus system to establish the concentration dependence of the effect. In addition, the phenomenon was investigated in a homologous system with custom-made HAV and B19V antibody-free and -containing human immunoglobulin intermediates. Viruses were analyzed with infectivity assays and fully validated polymerase chain reaction assays that also circumvent the obscuring effects of neutralizing antibodies with infectivity assays. RESULTS: By use of the heterologous mice minute virus and the homologous HAV and B19V systems, viruses passed the 35-nm (Planova 35N) filter in the absence of specific antibodies. Beyond a threshold virus antibody concentration, nanofiltration resulted in effective virus removal of viruses smaller than the nominal pore size of the filter used. CONCLUSION: HAV and B19V are effectively removed by antibody-dependent 35N nanofiltration, already at intermediate antibody concentrations well below those comparable to human plasma pools for fractionation.