Sp. Richieri et al., CHARACTERIZATION OF HIGHLY PURIFIED, INACTIVATED HIV-1 PARTICLES ISOLATED BY ANION-EXCHANGE CHROMATOGRAPHY, Vaccine, 16(2-3), 1998, pp. 119-129
Citations number
26
Categorie Soggetti
Veterinary Sciences",Immunology,"Medicine, Research & Experimental
This report characterizes inactivated, gp120 depleted, HIV-1 particles
purified by an anion exchange chromatography production process. This
antigen formulated with incomplete Freund's adjuvant constitutes Remu
ne(TM), which is being evaluated in a phase III clinical endpoint tria
l to determine the effect of this immune-based therapy on clinical pro
gression of HIV-1 seropositive patients. Multiple production lots of t
he inactivated HN-I antigen strain HZ321, isolated by anion exchange c
hromatography, exhibit purity of > 95% by gel filtration. These findin
gs are corroborated by thin section electron microscopy showing a homo
genous field of intact particles. Analyses of the purified virus par-t
ides for protein, lipid, carbohydrate and RNA show structural retentio
n of the envelope proteins, lipid bilayer and core components after la
rge scale processing. The qualitative identification of at least 85% o
f total HIV-1 protein is determined by ELISA, Western blot HPLC and am
ino acid sequencing analyses. Quantitative values are assigned to 50%
of these proteins. The data confirm the presence of virally encoded pr
oteins p6, p7, p(1)15, p17, p24, p32, p(1)39(Gag), gp41, p(p)55(Gag),
p66/51, Vpr; Vif and Nef: Excellent consistency between production lot
s and equivalency to HN-I preparations purified by sucrose density gra
dient sedimentation has been established for protein and lipid composi
tion, and overall purity. These findings further establish that non-vi
ral encoded proteins and lipids are integral structural components of
the intact virion and are not contaminants unique to a particular isol
ation method. The data confirm the presence of multicomponent antigens
in the viral particles for stimulating a broad HIV-1 specific immune
response. Finally, the work demonstrates that the two inactivation pro
cedures (beta-propiolactone and gamma irradiation), which achieve effi
cient viral inactivation meeting US FDA guidelines, do not damage the
protein antigens of the viral particles. (C) 1997 Elsevier Science Ltd
. All rights reserved.