Quantification of recombinant core-like particles of bluetongue virus using immunosorbent electron microscopy.
Immunosorbent electron microscopy was used to quantify recombinant baculovirus-generated bluetongue virus (BTV) core-like particles (CLP) in either purified preparations or lysates of recombinant baculovirus-infected cells. The capture antibody was an anti-BTV VP7 monoclonal antibody. The CLP concentration in purified preparations was determined to be 6.6 x 10(15) particles/l. CLP concentration in lysates of recombinant baculovirus-infected cells was determined at various times post-infection and shown to reach a value of 3 x 10(15) particles/l of culture medium at 96 h post-infection. The results indicated that immunosorbent electron microscopy, aided by an improved particle counting method, is a simple, rapid and accurate technique for the quantification of virus and virus-like particles produced in large scale in vitro systems.
Zheng
Y Z
YZ
Department of Chemical Engineering, The University of Queensland, Australia. yuanz@qimr.edu.au
Hyatt
A
A
Wang
L F
LF
Eaton
B T
BT
Greenfield
P F
PF
Reid
S
S
D000818:Animals; D001820:Bluetongue virus / Q000235:genetics / Q000648:ultrastructure*; D002460:Cell Line; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D018411:Spodoptera / Q000166:cytology; D014758:Viral Core Proteins / Q000032:analysis* / Q000235:genetics
D011993:Recombinant Fusion Proteins; C074954:VP3 protein, Bluetongue virus; D014758:Viral Core Proteins; C066862:VP7 protein, orbivirus