PDF《Nagata》

1 A recombinant humanized monoclonal antibody completely protects mice against lethal challenge with Venezuelan equine encephalitis virus Wei-Gang Hu a *, Amanda L.

Phelps b , Scott Jager a , Damon Chau a , Charles C. Hu c , Lyn M. O'Brien b , Stuart D. Perkins b , Amanda J. Gates b , Robert J. Phillpotts b , and Les P. Nagata a a Defence Research and Development Canada - Suffield, Box 4000, Station Main, Medicine Hat, Alberta, Canada T1A 8K6 b Defence Science and Technology Laboratory, Porton Down, Salisbury, United Kingdom, SP4 0JQ c Faculty of Science, University of Waterloo,

200 University Avenue West, Waterloo, Ontario, Canada N2L 3G1 * Corresponding author: [email protected] (NON-CONTROLLED GOODS) DMC A REVIEW: GCEC December

2013 2 ABSTRACT A recombinant humanized antibody to Venezuelan equine encephalitis virus (VEEV) was constructed in a monocistronic adenoviral expression vector with a foot-and-mouth-disease virus-derived 2A self-cleavage oligopeptide inserted between the antibody heavy and light chains. After expression in mammalian cells, the heavy and light chains of the humanized antibody (hu1A4A1IgG1-2A) were completely cleaved and properly dimerized. The purified hu1A4A1IgG1-2A retained VEEV binding affinity and neutralizing activity similar to its parental murine antibody. The half-life of hu1A4A1IgG1-2A in mice was approximately

2 days. Passive immunization of hu1A4A1IgG1-2A in mice (50 ?g/mouse)

24 hr before or after virulent VEEV challenge provided complete protection, indicating that hu1A4A1IgG1-2A has potent prophylactic and therapeutic effects against VEEV infection. Key Words: Recombinant humanized antibody, Venezuelan equine encephalitis virus, Prophylactic and therapeutic effects Running Title: Humanized antibody completely protects mice from lethal challenge with VEEV

3 1. Introduction Venezuelan equine encephalitis virus (VEEV), a single-stranded and positive-sense RNA alphavirus, is an important mosquito-borne pathogen in humans and equines [1]. VEEV causes a spectrum of human disease which includes acute encephalitis. Neurological disease appears in ~5 % of cases, and mortality is