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

3. Detection of B19 capsid proteins, VP-1 and VP-2,
in each eluted fraction by immuno-blot analysis
　1　　　2　　　　3　　　　4　　　　　5　　　　6
　7　　　8　　　9　　　10　　11　　　12　　13　　　14　　　　15
　　　　　　　　　　　　　　　　　　　　　　　　　peak 2
　16　　17　　18　　19　　　20　　　21　positive　
　　　　　　　　　　　　peak 3　　　　　 　 control
B19 spiked
solution
Pass through
peak 1

4. Characteristics and expected shape of B19 separated by Q-Sepharose
peak １ peak 2 peak 3
Capsid protein　　　 ○ 　　　　　　○ 　　　　　　×
DNase treatment resistant 　　sensitive 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 　　　 1.9　　 %
Pass through　　　　 ＜0.0* 　　　　　
Wash ＜0.0*
Eluted at
450 mM NaCl pH 5.8　 ＜0.0* 　 %
600 mM NaCl pH %
* 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