Improved stability of recombinant hemagglutinin using a formulation containing sodium thioglycolate

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• 作者：David G. Rhodes ; ^{drhodes@proteinsciences.com" class="auth_mail" title="E-mail the corresponding author} ; Kathy Holtz ; Pam Robinson ; Keyang Wang ; Clifton E. McPherson ; Manon M.J. Cox ; Indresh K. Srivastava ; ^{isrivastava@proteinsciences.com" class="auth_mail" title="E-mail the corresponding author}
• 关键词：rHA ; recombinant hemagglutinin ; PSC ; Protein Sciences Corporation ; BDS ; bulk drug substance ; SRID ; single radial immunodiffusion ; SDS&ndash ; PAGE ; sodium dodecyl sulfate&ndash ; polyacrylamide gel electrophoresis ; HAI ; hemagglutination inhibition
• 刊名：Vaccine
• 出版年：2015
• 出版时间：4 November 2015
• 年：2015
• 卷：33
• 期：44
• 页码：6011-6016
• 全文大小：716 K

文摘

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.