Improved stability of recombinant hemagglutinin using a formulation containing sodium thioglycolate
| Autor: | Keyang Wang, Clifton McPherson, Pam Robinson, Kathy Holtz, Indresh K. Srivastava, David G. Rhodes, Manon M.J. Cox |
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| Rok vydání: | 2015 |
| Předmět: |
Immunodiffusion
Antibodies Viral law.invention chemistry.chemical_compound law Immunology and Microbiology(all) Sodium citrate Potency Animals Polyacrylamide gel electrophoresis Vaccine Potency Radial immunodiffusion Mice Inbred BALB C Chromatography General Veterinary General Immunology and Microbiology Immunogenicity Public Health Environmental and Occupational Health Hemagglutinin veterinary(all) SODIUM THIOGLYCOLATE Dynamic Light Scattering Recombinant Proteins Infectious Diseases Hemagglutinins chemistry Biochemistry Influenza Vaccines Thioglycolates Recombinant DNA Molecular Medicine Electrophoresis Polyacrylamide Gel |
| Zdroj: | Vaccine. 33(44) |
| ISSN: | 1873-2518 |
| Popis: | This study was designed to improve the stability of liquid formulations of recombinant influenza hemagglutinin (rHA) and to understand the mechanism of early loss of potency for rHA. The potency of rHA derived from several influenza strains was determined using single radial immunodiffusion (SRID), and the structure of the rHA was characterized using SDS-PAGE and dynamic light scattering. rHA formed disulfide cross-linked multimers, and potency decreased during extended storage. To reduce disulfide-mediated cross-linking and early potency loss, rHA was formulated with sodium thioglycolate (STG) and citrate. Addition of 80 mM STG and 55 mM sodium citrate inhibited disulfide-mediated cross-linking without affecting protein function for each rHA tested. The shelf life of the rHA formulation with STG-citrate, based on potency as determined by SRID, was extended as much as 20-fold, compared to a control formulation without STG-citrate. STG-citrate did not have a significant effect on the immunogenicity of H1 A/California/7/2009 rHA in mice. |
| Databáze: | OpenAIRE |
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