Perfusion chromatography is a technique that arises to overcome the problem associated with mass transfer in the separation of large molecules such as proteins by liquid chromatography. Product production began as early as 1990s. Perfusion media are based on high crosslinked polystyrene divinylbenzene (PS-DVB) with two sets of pores: throughpores (6000-8000 Å) and diffusive pores (800-1500 Å). Throughpores enable better access of macromolecules to the inner of the particle, and increase the mobile phase velocity without any reduction of the column separation. With a structure of rich diffusion holes, the specific surface area of perfusion media is not reduced due to the presence of thoughpores. The dynamic capability of the perfusion chromatography media is stable with the increase of the flow rate.
Perfusion chromatography is an ideal choice the separation and purification of biological macromolecules widely used in ion-exchange, affinity, and hydrophobic interactive chromatography. They have excellent performance for complex biological samples with high viscosity, large molecule, high mass transfer barrier, and unstable biomolecules. The safety and reliability of perfusion chromatography technology have been proven in the production of FDA-approved biotherapeutic products.
Tradition Liquid Chromatography Media
GALAK Perfusion Chromatography Media
Advantages of Perfusion Chromatography in Enveloped Virus Vaccine
1. One-step purification for enveloped virus
Simulating the affinity between sulfated polysaccharides and enveloped virus to achieve the separation and purification for the virus.
The purpose of concentration and purification are all completed during perfusion chromatography process.
The time of the purification process for one batch virus vaccine will be reduced by 25-50%.
2. High yield, High purity, High removal rate of heteroprotein
The recovery rate of virus in finished virus vaccine can be increased by 15-35%, because of the removal of ultrafiltration membrane filtration process. The design of extra-large through pores and diffusive pores with " soft tentacle" ligand in perfusion chromatography media can protect the structure of virus protein for the damage of shear force and steric hindrance.
The affinity between virus protein and chromatography media can directly separate virus protein with impurities during the chromatography method. No need for extra process.
3. Long lifetime and low usage amount for chromatography media
The hydrophilic PS-DVB substrate has much higher mechanical strength than "soft" medium, which has higher pressure and longer repeated use time. The reused time can increase by 25-85%.
With the special design of GALAK perfusion media, the usage amount of media can decrease by 25-98%, compare with size exclusion chromatography.
4. Low cost for equipment and maintenance
GALAK purfusion chromatography just needs normal industrial liquid chromatography equipment.
No organic solvent is added during whole purification process.
Mild elution conditions, no harm for virus protein and media. Multiple CIP conditions meet different purification application.
Simple pretreatment for mixed virus solution, solid-liquid separation to remove visible impurities.
5. Application Range
|Rabies||Feline Calicivirus||Herpes Simplex gA and gB Glycoprotein Subunits|
|Influenza||Respiratory Syncytial Virus||Hepatitis B Surface Antigen|
|Japanese Enchephalitis||Human Herpes Simplex||Filamentous Hemagglutinin from B. pertussis|
|Feline Leukemia||Human Measles||Leucocytosis Promoting Factor Hemagglutinin|
|Feline Herpes||Human Parainfluenza|
Inapplicable condition for existing perfusion chromatography media
1. Non-envelope virus.
2. Inactive virus with formaldehyde (HCOH).
3. Non-suspension culture for the virus.
4. The protein size is too big or too small.