4.5 Article

Structural Stability of Hepatitis C Virus Envelope Glycoprotein E1: Effect of pH and Dissociative Detergents

Journal

JOURNAL OF PHARMACEUTICAL SCIENCES
Volume 98, Issue 9, Pages 3340-3357

Publisher

ELSEVIER SCIENCE INC
DOI: 10.1002/jps.21657

Keywords

hepatitis C virus; E1; protein particle; stability; empirical phase diagram

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The hepatitis C virus (HCV) envelope glycoprotein El has been widely employed as a potential vaccine antigen in clinical research. A truncated form (amino acids 192-326) of the El protein (Ely) was expressed in the yeast Hansenula polymorpha and purified from the cell lysate. Ely forms protein particles in the absence of detergent and remains monomeric when detergent concentration is high. In this work, a variety of spectroscopic and hydrodynamic techniques including circular dichroism, intrinsic and ANS fluorescence as well as static and dynamic light scattering are employed to evaluate Ely structural stability. The effect of two dissociative detergents, Empigen BB and Zwittergent 3-12 on Ely stability, is investigated and the results are summarized using the empirical phase diagram (EPD)-based approach. The EPDs reveal that when temperature is increased, Ely particles are more thermally stable than monomers at both pH 5 and 7. A more detailed biophysical characterization of the Ely particles is also performed including pH and temperature as variables. The EPD indicates that Ely particles are most stable at pH 7 and 8 under the given experimental conditions. The results from this study provide detailed information that will help guide the future development of El-based HCV vaccines. (C) 2008 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 98:3340-3357, 2009

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