Disruption of leaves and initial extraction of wildtype CPMV virus as a basis for producing vaccines from plants.
Wildtype cowpea mosaic virus (CPMV) was extracted from fresh and frozen plant material by methods suitable for large-scale application. Deep freezing, crushing, and thawing in water or buffers gave 0.6+/-0.2 mg g(-1) of virus after up to 24 h. Release from sliced fresh leaves was lower at 0.14+/-0.03 mg g(-1). Homogenisation of frozen leaves for 1 min increased yield to a maximum, on average of 3.5 mg g(-1) but varying between batches from 2.2 to 4.8 mg g(-1) virus Long term storage at -80 degrees C increased subsequent yield by 2 mg g(-1) per year on average; the maximum was 10.4+/-1.9 mg g(-1) (665 days storage). Within a batch, similar yields were obtained between individual fresh plants, and from frozen versus fresh leaves. After homogenisation for 1 min, 90% of debris particles were smaller than 12 microm, half under 5 microm and 10% less than 1 microm. Homogenate (4% dry weight) was rheologically complex, exhibiting shear thinning with hysteresis at low shear rates which bears on subsequent processing. At shear rates above 200 s(-1), its apparent viscosity was 0.02 N s m(-2).
Nichols
M E K
ME
Department of Biochemical Engineering, The Advanced Centre for Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, UK.
Stanislaus
T
T
Keshavarz-Moore
E
E
Young
H A
HA
D001709:Biotechnology; D017798:Comovirus / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D007887:Fabaceae / Q000821:virology*; D005615:Freezing; D010316:Particle Size; D018515:Plant Leaves / Q000821:virology; D012212:Rheology; D014614:Vaccines, Synthetic / Q000302:isolation & purification*; D014783:Viscosity
D014614:Vaccines, Synthetic