Size and conformational stability of the hepatitis A virus used to prepare VAQTA, a highly purified inactivated vaccine

Journal of Pharmaceutical Sciences
D B VolkinC Russell Middaugh

Abstract

A variety of biophysical techniques have been employed to examine the size and conformational integrity of highly purified hepatitis A virus (HAV) in solution (purified HAV particles are subsequently formalin-inactivated and adsorbed to aluminum salts for use as the vaccine VAQTA). The size of HAV particles was assessed by a combination of electron microscopy, sedimentation velocity, and dynamic light scattering. The effect of ionic strength and temperature on the overall conformational stability of HAV was determined by a combination of intrinsic HAV protein fluorescence, fluorescent probes of both RNA and protein, and UV-visible spectroscopy. A major structural change in HAV occurs near 60 degrees C with the addition of 0.2 M magnesium chloride enhancing the thermal stability of HAV by approximately 10 degrees C. Salt concentrations above 0.2 M, however, decrease the solubility of HAV. The effect of pH on the physical properties of HAV particles was monitored by dynamic light scattering, analytical size exclusion HPLC, and interaction with fluorescent dyes. HAV particles undergo a substantially reversible association/aggregation at pH values below 6 with the concomitant exposure of previously buried hydrophobic surfaces below...Continue Reading

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Citations

Apr 30, 2013·Food and Environmental Virology·Christina JarkeThiemo Albert
Aug 30, 2002·Seminars in Pediatric Infectious Diseases·Beth P Bell
Oct 27, 2009·Applied and Environmental Microbiology·Helen RawsthorneLee-Ann Jaykus
Jan 18, 2003·Journal of Pharmaceutical Sciences·Duane T BrandauC Russell Middaugh
Nov 28, 2013·Journal of Pharmaceutical Sciences·Wei QiC Russell Middaugh
Apr 8, 2009·International Journal of Food Microbiology·L BaertM Uyttendaele
Jun 25, 2015·Journal of Applied Microbiology·S E Arthur, K E Gibson
Jan 15, 2014·International Journal of Food Microbiology·Hayriye BozkurtDoris H D'Souza
Jul 6, 2014·Biologicals : Journal of the International Association of Biological Standardization·Ozan S KumruDavid B Volkin
Dec 21, 2014·Food and Environmental Virology·Sabastine E Arthur, Kristen E Gibson
Jul 30, 2015·Journal of Food Protection·Hayriye BozkurtP Michael Davidson
Mar 19, 2014·Applied and Environmental Microbiology·Hayriye BozkurtP Michael Davidson
Sep 16, 2020·Food and Environmental Virology·Waled M El-SenousyIbrahim I Alhawary
Mar 31, 1998·Journal of Pharmaceutical Sciences·C R MiddaughD R Casimiro

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