title: Influence of protein formulation and carrier solution on asymmetrical flow field-flow fractionation: a case study of the plant-produced recombinant anthrax protective antigen pp-PA83.
abstract: Asymmetrical flow field-flow fractionation (afFFF) was used to investigate the properties of a plant-produced anthrax toxin protective antigen, pp-PA83. The afFFF fractogram consisted of two main peaks with molar masses similar to the molecular mass of pp-PA83 monomer. afFFF carrier solutions strongly influenced the ratio and the intensity of the two main peaks. These differences indicate that conformation changes in the pp-PA83 molecule occurred during the afFFF analysis. Similar fractograms were obtained for different pp-PA83 formulations when the afFFF carrier solution and the protein formulation were the same (or very similar). The data show that in specific cases, afFFF could be used to study protein conformation and document the importance of studying the influence of the carrier solution on afFFF.
authors.affiliation: Therapeomic Inc., BioPark Rosental, Mattenstrasse 22, Basel, 4002, Switzerland.
authors.forename: Caroline
authors.initials: C
authors.lastname: Palais
authors.forename: Jessica A
authors.initials: JA
authors.lastname: Chichester
authors.forename: Slobodanka
authors.initials: S
authors.lastname: Manceva
authors.forename: Vidadi
authors.initials: V
authors.lastname: Yusibov
authors.forename: Tudor
authors.initials: T
authors.lastname: Arvinte
mesh_terms: D000942:Antigens, Bacterial / Q000737:chemistry* / Q000378:metabolism; D001427:Bacterial Toxins / Q000737:chemistry* / Q000378:metabolism; D002626:Chemistry, Pharmaceutical; D031521:Fractionation, Field Flow / Q000379:methods*; D019999:Pharmaceutical Solutions / Q000737:chemistry* / Q000378:metabolism
chemical_list: D000942:Antigens, Bacterial; D001427:Bacterial Toxins; D019999:Pharmaceutical Solutions; C030325:anthrax toxin
keywords: analysis; anthrax protective antigen; carrier solution; flow field-flow fractionation; protein aggregation; protein conformation; protein formulation; protein structure; vaccines