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E. &. O. O. &. Purification of viruses. Purification. To obtain the viral particles in a single form out of cell debris i.e. removal of all impurities. Purification outline. Preparation of harvest. Methods of purification. Criteria of purity. Maintenance of purified virus.
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Purification To obtain the viral particles in a single form out of cell debris i.e. removal of all impurities.
Purification outline • Preparation of harvest. • Methods of purification. • Criteria of purity. • Maintenance of purified virus.
Preparation of harvest Bacteria Insect Animal Plant
Methods of purification • Filtration. • Centrifugation. • Adsorption. • Precipitation. • Solvent extraction. • Other methods.
Methods of purification • Filtration. • Centrifugation. • Adsorption. • Precipitation. • Solvent extraction. • Other methods.
Filtration : 1.Ultrafiltration 2. Gel filtration
Methods of purification • Filtration. • Centrifugation. • Adsorption. • Precipitation. • Solvent extraction. • Other methods.
centrifugation Differential Cushion Isopycnic Gradient
Methods of purification • Filtration. • Centrifugation. • Adsorption. • Precipitation. • Solvent extraction. • Other methods.
Adsorption : 1.Ion exchange 2. Heamadsroption
Methods of purification • Filtration. • Centrifugation. • Adsorption. • Precipitation. • Solvent extraction. • Other methods.
Precipitation : 1. Chemical • Ammoniumsufate • PEG • Zinc acetae 2. Physical • Isoelectric piont (pH change)
Methods of purification • Filtration. • Centrifugation. • Adsorption. • Precipitation. • Solvent extraction. • Other methods.
Methods of purification • Filtration. • Centrifugation. • Adsorption. • Precipitation. • Solvent extraction. • Other methods.
Other methods • Electrophoresis. • Serological methods. • Enzymatic reactions. • HPLC
Criteria of purity Chemical purity Physical purity Biological purity
Criteria of purity E.M. Spectrophotometer serological Isoradiation Infectivity
Criteria of purity E.M. Spectrophotometer serological Isoradiation Infectivity
Criteria of purity E.M. Spectrophotometer serological Isoradiation Infectivity
Infectivity: 1. Local lesion assay 2. Plaque assay
Criteria of purity E.M. Spectrophotometer serological Isoradiation Infectivity
Criteria of purity E.M. Spectrophotometer serological Isoradiation Infectivity
4- Maintenance: The half life of infectivity can be measured in seconds at 60 oC; minutes at 37 oC, hours at 20oC , days at 4 oC and years at -70 oC or lower,(Fenner et al, 1987) Lyophlization