title: Analysis of the thermal and pH stability of human respiratory syncytial virus.
abstract: Respiratory syncytial virus (RSV) was studied as a function of pH (3-8) and temperature (10-85 degrees C) by fluorescence, circular dichroism, and high-resolution second-derivative absorbance spectroscopies, as well as dynamic light scattering and optical density as a measurement of viral aggregation. The results indicate that the secondary, tertiary, and quaternary structures of RSV are both pH and temperature labile. Derivative ultraviolet absorbance and fluorescence spectroscopy (intrinsic and extrinsic) analyses suggest that the stability of tertiary structure of RSV proteins is maximized near neutral pH. In agreement with these results, the secondary structure of RSV polypeptides seems to be more stable at pH 7-8, as evaluated by circular dichroism spectroscopy. The integrity of the viral particles studied by turbidity and dynamic light scattering also revealed that RSV is more thermally stable near neutral pH and particularly prone to aggregation below pH 6. By combination of the spectroscopic data employing a multidimensional eigenvector phase space approach, an empirical phase diagram for RSV was constructed. The pharmaceutical utility of this approach and the optimal formulation conditions are discussed.
authors.affiliation: Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, Kansas 66047, USA.
authors.forename: Salvador F
authors.initials: SF
authors.lastname: Ausar
authors.forename: Jason
authors.initials: J
authors.lastname: Rexroad
authors.forename: Vladimir G
authors.initials: VG
authors.lastname: Frolov
authors.forename: Jee L
authors.initials: JL
authors.lastname: Look
authors.forename: Nandini
authors.initials: N
authors.lastname: Konar
authors.forename: C Russell
authors.initials: CR
authors.lastname: Middaugh
mesh_terms: D002942:Circular Dichroism; D006358:Hot Temperature; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008027:Light; D018113:Respiratory Syncytial Virus, Human / Q000254:growth & development / Q000502:physiology* / Q000648:ultrastructure; D012542:Scattering, Radiation; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry; D013816:Thermodynamics