Purification To obtain the viral particles in a

Purification To obtain the viral particles in a single form out of cell debris i. e. removal of all impurities.

Purification outline 1. Preparation of harvest. 2. Methods of purification. 3. Criteria of purity. 4. Maintenance of purified virus.

Preparation of harvest Bacteria Animal Plant Insect

Methods of purification 1. Filtration. 2. Centrifugation. 3. Adsorption. 4. Precipitation. 5. Solvent extraction. 6. Other methods.

Methods of purification 1. Filtration. 2. Centrifugation. 3. Adsorption. 4. Precipitation. 5. Solvent extraction. 6. Other methods.

Filtration : 1. Ultrafiltration 2. Gel filtration

Ultra-filtration

Gel filtration

Methods of purification 1. Filtration. 2. Centrifugation. 3. Adsorption. 4. Precipitation. 5. Solvent extraction. 6. Other methods.

centrifugation Differential Isopycnic Gradient Cushion

Methods of purification 1. Filtration. 2. Centrifugation. 3. Adsorption. 4. Precipitation. 5. Solvent extraction. 6. Other methods.

Adsorption : 1. Ion exchange 2. Heamadsroption

Ion exchange chromatography

Heamadsorption

Methods of purification 1. Filtration. 2. Centrifugation. 3. Adsorption. 4. Precipitation. 5. Solvent extraction. 6. Other methods.

Precipitation : 1. Chemical • Ammoniumsufate • PEG • Zinc acetae 2. Physical • Isoelectric piont (p. H change)

Methods of purification 1. Filtration. 2. Centrifugation. 3. Adsorption. 4. Precipitation. 5. Solvent extraction. 6. Other methods.

Methods of purification 1. Filtration. 2. Centrifugation. 3. Adsorption. 4. Precipitation. 5. Solvent extraction. 6. Other methods.

Other methods • • Electrophoresis. Serological methods. Enzymatic reactions. HPLC

Electrophoresis

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

Marking

Criteria of purity E. M. Spectrophotometer serological Isoradiation Infectivity

Infectivity: 1. Local lesion assay 2. Plaque assay

Virus preparation

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 o. C; minutes at 37 o. C, hours at 20 o. C , days at 4 o. C and years at -70 o. C or lower, (Fenner et al, 1987) Lyophlization