Parvovirus B19 Inactivation Results obtained at the Paul-Ehrlich-Institute

1. PEI Parvovirus B19 Inactivation Results obtained at the Paul-Ehrlich-Institute Johannes Blümel Head of Viral Safety Section Paul-Ehrlich-Institut Federal Agency for Sera and Vaccines Langen, Germany

2. 350 2659 PEI B19V Genome Structure and Gene Expression IM-81: 131bp Insert P6 Promoter DNA ITR ITR NS1 mRNA NS1 Protein VP1 mRNA 350 406 3045 5010 2030/ 1910 VP1 Protein VP2 mRNA 350 406 2183 3045 5010 2030/ 1910 VP2 Protein 350 406 2183 4704 5010 2030/ 1910

3. PEI Kinetics of Capsid mRNA in Ku812Ep6 cells Blümel et al., J. Virol 79, 14197-14206 (2005)

4. PEI Capsid Antigen Expression in KU812Ep6-Cells Genotype 1 Genotype 2 2 12 24 36 48 72 2 12 24 36 48 72 h p.i. kDa 185 98 53 Blümel et al., J. Virol 79, 14197-14206 (2005)

5. 0 200 100 300 400 500 PEI Capsid Protein Variations B19V genotypes 1 and 2 VP2 VP1 700 600 Position Type 1 Type 2 4 K E 18 A D 21 Q K 68 N S 72 N D 96 S P 98 N S 102 A T 123 V I 170 H V 192 V A 209 F L 247 S P 259 S T 370 D T 372 T S 531 S T 533 S N 616 M I 447 Q E 454 T S 550 N S Do these variations influence stability of capsids?(thermosensitivity, low pH) Blümel et al., J. Virol 79, 14197-14206 (2005)

6. PEI Thermal inactivation occurs by disintegration of capsids - DNAse + DNAse Heat 55 56 60 60 RT 52 53 54 DNAse I, 1h, 37°C DNA Purification Southern Analysis Blümel et al. 2002, Transfusion 42:1011-1018

7. PEI Inactivation of Type 1 versus Type 2 (in 5% albumin at 56°C) Blümel et al., J. Virol 79, 14197-14206 (2005)

8. PEI Inactivation of Type I versus Type II (in 5% albumin, at pH4, 37°C) Blümel et al., J. Virol 79, 14197-14206 (2005)

9. PEI c-ETS SP1 YY1 c-ETS Direct Repeat Octamer YY1 SP1 Direct Repeat E4BP4 YY1 SP1 SP1/MZF1 TATA box [Blümel et al., J. Virol, 2005]