Chromatographic removal combined with heat, acid and chaotropic inactivation of four model viruses.
The virus removal of protein A affinity chromatography, inactivation capacity, acid pH and a combination of high temperature with a chaotropic agent was determined in this work. The model viruses studied were sendaivirus, human immunodeficency virus (HIV-IIIb), human poliovirus type-II, human herpesvirus I and canine parvovirus. The protein A affinity chromatography showed a maximum reduction factor of 8 logs in the case of viruses larger than 120 nm size, while for small viruses (18-30 nm) the maximum reduction factor was about 5 logs. Non viral inactivation was observed during the monoclonal antibody elution step. Low pH treatment showed a maximum inactivation factor of 7.1 logs for enveloped viruses. However, a weak inactivation factor (3.4 logs) was obtained for DNA nonenveloped viruses. The combination of high temperature with 3 M KSCN showed a high inactivation factor for all of the viruses studied. The total clearance factor was 23.1, 15.1, 13.6, 20.0 and 16.0 logs for sendaivirus, HIV-IIIb, human poliovirus type-II, human herpesvirus I and canine parvovirus, respectively.
Valdés
R
R
Center for Genetic Engineering and Biotechnology, Ave 31/158 and 190, PO Box 6162, Havana 10600, Cuba.
Ibarra
Neysi
N
Ruibal
I
I
Beldarraín
A
A
Noa
E
E
Herrera
N
N
Alemán
R
R
Padilla
S
S
Garcia
J
J
Pérez
M
M
Morales
R
R
Chong
E
E
Reyes
B
B
Quiñones
Y
Y
Agraz
A
A
Herrera
L
L
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis; D002846:Chromatography, Affinity / Q000379:methods*; D004285:Dogs; D004340:Drug Contamination / Q000517:prevention & control; D005240:Feasibility Studies; D006678:HIV / Q000276:immunology / Q000302:isolation & purification; D006514:Hepatitis B Surface Antigens / Q000276:immunology*; D017325:Hepatitis B Vaccines / Q000096:biosynthesis; D006358:Hot Temperature; D006801:Humans; D006863:Hydrogen-Ion Concentration; D051379:Mice; D008807:Mice, Inbred BALB C; D017992:Parvovirus / Q000276:immunology / Q000302:isolation & purification; D017955:Poliovirus / Q000276:immunology / Q000302:isolation & purification; D029082:Sendai virus / Q000276:immunology / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D013205:Staphylococcal Protein A / Q000737:chemistry / Q000276:immunology; D015164:Vaccines, Inactivated; D014614:Vaccines, Synthetic; D014780:Viruses / Q000276:immunology / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal; D006514:Hepatitis B Surface Antigens; D017325:Hepatitis B Vaccines; D013205:Staphylococcal Protein A; D015164:Vaccines, Inactivated; D014614:Vaccines, Synthetic