Title
High-throughput screening of stabilizers for Respiratory Syncytial Virus: Identification of stabilizers and their effects on the conformational thermostability of viral particles
Year
2007
Journal
Human Vaccines
Abstract
Respiratory syncytial virus (RSV) is the leading cause of severe respiratory infection inchildren worldwide. Recombinant live attenuated viral preparations are one of the most promisingstrategies for vaccination but they typically possess poor thermostability. In this work, a library ofcompounds was screened and stabilizers were selected based on their ability to inhibit theaggregation of RSV perturbed at 56 °C. After screening and selection of excipients, theconformational stability of the RSV proteins was evaluated in the presence of potential stabilizers.The secondary and tertiary structures as well as aggregation/dissociation of RSV were monitoredusing circular dichroism and second derivative UV absorption spectroscopies and light scattering,respectively, as a function of temperature (10-90 °C). RSV membrane fluidity was also evaluatedby generalized polarization of Laurdan fluorescence. Screening experiments showed that a varietyof sugars, amino acids, polyols and polyanions inhibited the aggregation of viral particles.Conformational stability studies demonstrated that the addition of sugars and polyols stabilizedRSV as indicated by a significant increase in the transition melting temperature (Tm) of both thesecondary and tertiary structures as well as the gel to liquid crystalline membrane transition. Theseresults should provide the basis for rational development of more physically stable formulations oflive attenuated RSV vaccines.
Instrument
J-810
Keywords
Circular dichroism, Secondary structure, Thermal stability, Chemical stability, Biochemistry, Corona Virus