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.

Pseudorabies PRV virus vaccine

Chromatography  Purification Test For Inactivated Pseudorabies Virus

Result

ComponentTotal Protein Concentration (mg/ml)Volume (mL)
Original Sample Diluent0.94730
Elution0.3515
Removing rate of impure protein94%

Chromatography Condition

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

Chromatography Process:

Column Balance → Sample Loading → Re-balance → Elution

Collect the conponents of flow-though process and Elution process

Analysis Methods:

(1) SEC-HPLC Analysis for every components

(2) BCA protein concentration test

chromatography chart for One-step chromatography Purification of Inactivated Pseudorabies PRV Virus Vaccine

Result Analysis

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.

Chromatography gram for the sample of Inactivated Pseudorabies PRV Virus
Chromatography gram for the elution liquid of Inactivated Pseudorabies Virus
Chromatography gram for the flow-though-liquid-of-Inactivated Pseudorabies Virus

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