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Japanese Red Cross Plasma Fractionation Center K. Furuya, T. Yokoyama, H. Maeno, T. Murozuka

Analysis of human parvovirus B19 components and strategies of non-enveloped virus removal from factor VIII concentrates. Japanese Red Cross Plasma Fractionation Center K. Furuya, T. Yokoyama, H. Maeno, T. Murozuka M. Tanifuji, A. Wakisaka and T. Tomono. NaCl [mM ]. 38 x 10 n IU / mL. pH 5.5.

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Japanese Red Cross Plasma Fractionation Center K. Furuya, T. Yokoyama, H. Maeno, T. Murozuka

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  1. Analysis of human parvovirus B19 components and strategies of non-enveloped virus removal from factor VIII concentrates Japanese Red Cross Plasma Fractionation Center K. Furuya, T. Yokoyama, H. Maeno, T. Murozuka M. Tanifuji, A. Wakisaka and T. Tomono

  2. NaCl [mM] 38 x 10n IU / mL pH 5.5 Pass through Peak 1 Peak 2 Peak 3 pH 6.4 Fraction number Elution profile for B19 components on Q-Sepharose

  3. Pass through peak 1 B19 spiked solution 123456 789101112131415 peak 2 161718192021positive peak 3control Detection of B19 capsid proteins, VP-1 and VP-2, in each eluted fraction by immuno-blot analysis

  4. Characteristics and expected shape of B19 separated by Q-Sepharose peak １ peak 2 peak 3 Capsid protein　　　 ○ 　　　　　　○ 　　　　　　× DNase treatment resistantsensitive sensitive Charge not negative negative negative Shape

  5. Elution profile for B19 on Q-Sepharose after nanofiltration NaCl [mM] 38 x 10n IU / mL pH 5.5 Not filtrated◆ 20 nm filtration ■ 35 nm filtration ▲ ◆ ▲ Pass through ▲ pH 6.4 ■ Fraction number

  6. B19 DNA fragment and factor VIII with different elution conditions B19 DNA fragment Amount of FVIII （38 x 10n IU/ mL） ( U ) Spiked solution 1.9100% Pass through　　　　 ＜0.0* Wash ＜0.0* Eluted at 450 mM NaCl pH 5.8＜0.0*97.6% 600 mM NaCl pH 5.5 1.7 4.2% * Not detected

  7. Conclusions 1. At least three forms of B19 were found ,i.e. , intact virus virion, disrupted virion and DNA fragment 2. B19 virus virion and disrupted virion can be removed by implementing a 20-nm-pore-size filter 3. B19 DNA fragment can be separated from factor VIII on Q-Sepharose with adequate elution conditions

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