One-step Purification For Inactivated Pseudorabies Virus Vaccine
Pseudorabies (Aujeszky's disease) is a viral disease caused by pseudorabies virus (PRV). PRV is mainly spread via direct contact, but may also transmit by air, water and contaminated fomites. Outbreaks of PRV in pigs are difficult to control, causing catastrophic economic losses in the swine industry. PRV vaccines are the most effective to control virus spread. Inactivated vaccine is one of the common PRV vaccine in the market. Traditional purification process (membrane filtration and size exclusion) has low virus protein recovery rate. GALAK provide new technology of VirCap® perfusion chromatography media for PRV virus.
GALAK perfusion chromatography media is based on high crosslinked polystyrene divinylbenzene (PS-DVB) with two sets of pores: throughpores (6000-8000 Å) and diffusive pores (800-1500 Å). The PS-DVB particles increase the mechanical strength of the substrate, improve the service life of VirCap®. The big pores let big protein molecules go though without break the protein structure. The unique affinity interaction between protein and VirCap® ligand makes the virus separation much easier.
Chromatography Purification Test For Inactivated Pseudorabies Virus
|Component||Total Protein Concentration (mg/ml)||Volume (mL)|
|Original Sample Diluent||0.947||30|
|Removing rate of impure protein||94%|
Column: VirCap AF® column (ID7.3mm*100ml), CV=4.2ml
Balance/Washing Buffer: 20mM MES pH 6.5-7.5, or 20mM PB pH 7.0-8.0 (buffer A)
Elution Buffer: 1M KCl in buffer A (buffer B)
Flow Rate: 0.5-1.0 ml/min
Detector: 280nm & 260nm
Sample: Original sample / 1-2 times dilution with buffer A
Column Balance → Sample Loading → Re-balance → Elution
Collect the conponents of flow-though process and Elution process
(1) SEC-HPLC Analysis for every components
(2) BCA protein concentration test
SEC-HPLC analysis was performed on the component of sample, break-though and elution. The result shows the target substance did not peak at RT=10min in break-though component. VirCap® AF media specifically adsorbed this component and eluted the virus particles by changing the ionic strength in the mobile phase.
The purity and concentration of the virus have been significantly improved.