title: Improved stability of recombinant hemagglutinin using a formulation containing sodium thioglycolate.
abstract: 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.
authors.affiliation: Protein Sciences Corporation, Meriden, CT 06450, United States. Electronic address: drhodes@proteinsciences.com.
authors.forename: David G
authors.initials: DG
authors.lastname: Rhodes
authors.affiliation: Protein Sciences Corporation, Meriden, CT 06450, United States.
authors.forename: Kathy
authors.initials: K
authors.lastname: Holtz
authors.affiliation: Protein Sciences Corporation, Meriden, CT 06450, United States.
authors.forename: Pam
authors.initials: P
authors.lastname: Robinson
authors.affiliation: Protein Sciences Corporation, Meriden, CT 06450, United States.
authors.forename: Keyang
authors.initials: K
authors.lastname: Wang
authors.affiliation: Protein Sciences Corporation, Meriden, CT 06450, United States.
authors.forename: Clifton E
authors.initials: CE
authors.lastname: McPherson
authors.affiliation: Protein Sciences Corporation, Meriden, CT 06450, United States.
authors.forename: Manon M J
authors.initials: MM
authors.lastname: Cox
authors.affiliation: Protein Sciences Corporation, Meriden, CT 06450, United States. Electronic address: isrivastava@proteinsciences.com.
authors.forename: Indresh K
authors.initials: IK
authors.lastname: Srivastava
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D000067493:Dynamic Light Scattering; D004591:Electrophoresis, Polyacrylamide Gel; D006388:Hemagglutinins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*; D005779:Immunodiffusion; D007252:Influenza Vaccines / Q000008:administration & dosage / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D013864:Thioglycolates / Q000737:chemistry*; D064166:Vaccine Potency
chemical_list: D000914:Antibodies, Viral; D006388:Hemagglutinins; D007252:Influenza Vaccines; D011994:Recombinant Proteins; D013864:Thioglycolates; C017487:2-mercaptoacetate
keywords: Influenza vaccine; SRID; STG; Stability; rHA