tetano
Editor, Senior Moderator
Vaccine. 2013 Jul 15. pii: S0264-410X(13)00933-X. doi: 10.1016/j.vaccine.2013.07.011. [Epub ahead of print]
Assessment of viral replication in eggs and HA protein yield of pre-pandemic H5N1 candidate vaccine viruses.
Jing X, Soto J, Gao Y, Phy K, Ye Z.
Source
Laboratory of Respiratory Viral Diseases, Division of Viral Products, Office of Vaccine Research and Review, United States Food and Drug Administration, 8800 Rockville Pike, Bethesda, MD 20892, United States.
Abstract
H5N1 infection and the potential for spread from human to human continue to pose a severe public health concern. Since vaccination remains the most effective way to prevent a potential H5N1 pandemic, the World Health Organization (WHO) Collaborating Centers (CCs) and Essential Regulatory Laboratories (ERLs) engineered and developed a panel of H5N1 pre-pandemic vaccine viruses for pandemic vaccine preparedness as well as production of antigen potency testing reagents (reference antigen and reference anti-serum) for vaccine standardization. To develop a strategy utilizing a number of biochemical methods for the characterization of the viral growth properties and protein yield in eggs, we have selected eight H5N1 pre-pandemic viruses and determined the viral Egg Infectious Dose 50 (EID50), total protein yield, hemagglutinin (HA) to nucleoprotein (NP) ratios (HA:NP), and HA1 content of each virus. Our results showed that all the tested H5N1 vaccine viruses grew to high titers in eggs. The total viral protein yield varies within a narrow range, whereas there were greater differences in the HA:NP protein ratios among the eight viruses. The RP-HPLC based HA1 content analysis demonstrated that the viruses A/Anhui/1/2010, A/Hubei/1/2005, and A/goose/Guiyang/337/2006 contained higher HA contents than other five viruses including A/Vietnam/1203/2003. Our approach for analyzing virus growth and protein yield will allow us identify optimal vaccine virus in a timely manner. In addition, we successfully purified the HA proteins of H5N1 vaccine viruses by optimizing bromelain cleavage conditions. Our studies on the HA protein purification may improve the quality control of the production of influenza vaccine test reagent.
Copyright ? 2013. Published by Elsevier Ltd.
KEYWORDS:
HA bromelain cleavage, HA purification, Influenza H5N1 virus protein yield, Influenza pandemic preparedness
PMID:
23867014
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/23867014
Assessment of viral replication in eggs and HA protein yield of pre-pandemic H5N1 candidate vaccine viruses.
Jing X, Soto J, Gao Y, Phy K, Ye Z.
Source
Laboratory of Respiratory Viral Diseases, Division of Viral Products, Office of Vaccine Research and Review, United States Food and Drug Administration, 8800 Rockville Pike, Bethesda, MD 20892, United States.
Abstract
H5N1 infection and the potential for spread from human to human continue to pose a severe public health concern. Since vaccination remains the most effective way to prevent a potential H5N1 pandemic, the World Health Organization (WHO) Collaborating Centers (CCs) and Essential Regulatory Laboratories (ERLs) engineered and developed a panel of H5N1 pre-pandemic vaccine viruses for pandemic vaccine preparedness as well as production of antigen potency testing reagents (reference antigen and reference anti-serum) for vaccine standardization. To develop a strategy utilizing a number of biochemical methods for the characterization of the viral growth properties and protein yield in eggs, we have selected eight H5N1 pre-pandemic viruses and determined the viral Egg Infectious Dose 50 (EID50), total protein yield, hemagglutinin (HA) to nucleoprotein (NP) ratios (HA:NP), and HA1 content of each virus. Our results showed that all the tested H5N1 vaccine viruses grew to high titers in eggs. The total viral protein yield varies within a narrow range, whereas there were greater differences in the HA:NP protein ratios among the eight viruses. The RP-HPLC based HA1 content analysis demonstrated that the viruses A/Anhui/1/2010, A/Hubei/1/2005, and A/goose/Guiyang/337/2006 contained higher HA contents than other five viruses including A/Vietnam/1203/2003. Our approach for analyzing virus growth and protein yield will allow us identify optimal vaccine virus in a timely manner. In addition, we successfully purified the HA proteins of H5N1 vaccine viruses by optimizing bromelain cleavage conditions. Our studies on the HA protein purification may improve the quality control of the production of influenza vaccine test reagent.
Copyright ? 2013. Published by Elsevier Ltd.
KEYWORDS:
HA bromelain cleavage, HA purification, Influenza H5N1 virus protein yield, Influenza pandemic preparedness
PMID:
23867014
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/23867014