Purification Techniques | Application of Gel Filtration Chromatography Resin in Viral Vaccines

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As a type of acellular organism, viruses typically parasitize living cells, utilizing the host's metabolic system to replicate and proliferate.

Biological virus is a non-cellular life form characterized by its small size and simple structure, usually containing only one type of nucleic acid, either DNA or RNA.

Although viruses often evoke fear, advancements in scientific research have enabled the use of partial or complete viral components, or processed viruses, to stimulate the human immune system to produce specific antibodies and immune cells. This has given rise to the concept of viral vaccines.

The development of viral vaccines undergoes rigorous scientific investigation and clinical trials to ensure their safety and efficacy. Common viral vaccines include influenza vaccines, rabies vaccines, chickenpox vaccines, hepatitis vaccines, and pneumonia vaccines.

Purification Process of Viral Vaccines

In the study of viruses, the purification process is an essential step. High-purity viral samples allow for more detailed research, and advanced separation and purification technologies play a critical role in the preparation of viral vaccines.

When it comes to vaccine purification methods, affinity chromatography, ion exchange chromatography, and hydrophobic chromatography may come to mind first. However, gel filtration chromatography is often overlooked. In this article, we will focus on the application of gel filtration chromatography resin in the purification of viral vaccines.

Gel Filtration Chromatography (GFC), also known as Size Exclusion Chromatography (SEC), separates samples based on molecular size by utilizing the pore size of spherical gel particles.

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Diagram of Gel Filtration Chromatography

Chromstar® 4FF, a representative gel filtration chromatography resin, is commonly used in the purification of recombinant proteins, antibodies, nucleic acids, viruses, virus-like particles, and polysaccharides. It has shown outstanding performance in influenza vaccines: on one hand, it can remove ovalbumin from egg-based influenza vaccines; on the other hand, it achieves excellent results in both HA yield and protein removal rate in cell-based influenza vaccines.

Technical Parameters

Name	Chromstar® 4FF
Chromatography Resin Type	Gel Filtration
Matrix	Cross-linked Agarose, 4%
Particle Size Range	45~165 μm
Separation Range	Spherical Molecules: 60 kDa~20,000 kDa Linear Molecules: 20 kDa~6,000 kDa
Recommended Flow Rate	90~200 cm/h*
Maximum Flow Rate	250 cm/h
Maximum Back Pressure	0.3 MPa
Working Temperature	4-30°C
pH Stability**	2-14
Compatible Solvents	Common aqueous solutions, 30% isopropanol, 75% ethanol, 1M NaOH, 1M acetic acid, 6M guanidine hydrochloride, 8M urea

*When the bed height is ≥ 60 cm, it is recommended to reduce the flow rate to 45-60 cm/h.

**The physicochemical properties and functionality of the chromatography resin remain stable after being placed at 40°C and pH 2-14 for 7 days.

Application Case

Purification of Influenza Virus Vaccine using Chromstar® 4FF

Resin: Chromstar® 4FF

Column: Chrom-LinX® 16/700 (H=60 cm)

Sample: Influenza virus

Sample Volume: 10 mL

Flow Rate: 6 mL/min

Buffer: 10 mM PBS

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Purification Cycle	HA Recovery Rate	Protein Removal Rate
First Cycle	96%	47%
Second Cycle	92%	48%
Third Cycle	91%	48%

Summary: After three rounds of purification using Chromstar® 4FF, the HA recovery rate consistently exceeded 90%, and the protein removal rate reached 47%.

Catalog Number

Bulk Packing Materials
Product Name	Catalog No.	Specification
Chromstar® 4FF	1010-1121	25 mL
	1010-1122	100 mL
	1010-1123	500 mL
	1010-1124	1L
	1010-1125	5L
	1010-1126	10L
	1010-1127	20 L

References

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13 Feb 2023