title: Biophysical and formulation studies of the Schistosoma mansoni TSP-2 extracellular domain recombinant protein, a lead vaccine candidate antigen for intestinal schistosomiasis.
abstract: A candidate vaccine to prevent human schistosomiasis is under development. The vaccine is comprised of a recombinant 9 kDa antigen protein corresponding to the large extracellular domain of a tetraspanin surface antigen protein of Schistosoma mansoni, Sm-TSP-2. Here, we describe the biophysical profile of the purified, recombinant Sm-TSP-2 produced in the yeast PichiaPink, which in preclinical studies in mice was shown to be an effective vaccine against intestinal schistosomiasis. Biophysical techniques including circular dichroism, intrinsic and extrinsic fluorescence and light scattering were employed to generate an empirical phase diagram, a color based map of the physical stability of the vaccine antigen over a wide range of temperatures and pH. From these studies a pH range of 6.0-8.0 was determined to be optimal for maintaining the stability and conformation of the protein at temperatures up to 25 °C. Sorbitol, sucrose and trehalose were selected as excipients that prevented physical degradation during storage. The studies described here provide guidance for maximizing the stability of soluble recombinant Sm-TSP-2 in preparation of its further development as a vaccine.
authors.affiliation: Macromolecule and Vaccine Stabilization Center; Department of Pharmaceutical Chemistry; University of Kansas; Lawrence, KS USA.
authors.forename: Weiqiang
authors.initials: W
authors.lastname: Cheng
authors.forename: Elena
authors.initials: E
authors.lastname: Curti
authors.forename: Wanderson C
authors.initials: WC
authors.lastname: Rezende
authors.forename: Clifford
authors.initials: C
authors.lastname: Kwityn
authors.forename: Bin
authors.initials: B
authors.lastname: Zhan
authors.forename: Portia
authors.initials: P
authors.lastname: Gillespie
authors.forename: Jordan
authors.initials: J
authors.lastname: Plieskatt
authors.forename: Sangeeta B
authors.initials: SB
authors.lastname: Joshi
authors.forename: David B
authors.initials: DB
authors.lastname: Volkin
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Hotez
authors.forename: C Russell
authors.initials: CR
authors.lastname: Middaugh
authors.forename: Maria Elena
authors.initials: ME
authors.lastname: Bottazzi
mesh_terms: D000818:Animals; D000947:Antigens, Helminth / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D055592:Biophysical Phenomena; D002942:Circular Dichroism; D004339:Drug Compounding; D004355:Drug Stability; D005079:Excipients / Q000494:pharmacology; D005470:Fluorometry; D015801:Helminth Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D006863:Hydrogen-Ion Concentration; D051379:Mice; D010843:Pichia / Q000235:genetics; D011487:Protein Conformation; D055550:Protein Stability; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D012555:Schistosomiasis mansoni / Q000276:immunology / Q000517:prevention & control*; D013696:Temperature; D059470:Tetraspanins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D014612:Vaccines / Q000737:chemistry* / Q000276:immunology* / Q000302:isolation & purification
chemical_list: D000947:Antigens, Helminth; D005079:Excipients; D015801:Helminth Proteins; D011994:Recombinant Proteins; C000589037:TSP-2 protein, Schistosoma mansoni; D059470:Tetraspanins; D014612:Vaccines