Process and Application of FVRCP Cat Vaccine

With the increasing number of pet cats in modern families, an unprecedentedly prosperous "cat economy" has emerged. According to data, the number of pet cats has increased to 65.36 million in 2022, and it is expected to continue growing in the future. At the same time, the prevention and control of cat infectious diseases have become particularly important.

Among them, feline calicivirus, feline viral rhinotracheitis, and feline panleukopenia are important infectious diseases of cats, which can cause respiratory and intestinal diseases in cats, resulting in lameness, abortion, chronic gastroenteritis, and even death in severe cases.

Vaccines are the best way to prevent these diseases, and there are already marketed products for vaccines against these three diseases.

The FVRCP vaccine typically refers to a trifunctional inactivated vaccine that can simultaneously prevent feline herpesvirus, feline calicivirus, and feline panleukopenia virus (feline distemper), meaning it's a single vaccine that can combat three viruses. One of the marketed products, Zoetis's Fel-O-vax PCT, requires four injections according to the recommended immunization schedule, which is detailed below.

The image is sourced from the internet.

Currently, both Chinese and foreign enterprises have made significant progress in the development of FVRCP vaccine, and below is an overview of its development progress. Among them, several animal vaccine companies have collaborated with BioLink in the downstream process development and manufacturing.

Company Nama	Product Name	Stage
Zoetis	Fel-O-Vax^® PCT	Launched in November 2022
Ringpu	RINVAC F RCP	Launched in January 2024
Bioally	Meow Beihu	Launched in February 2024
Yian	Miao San Xin	Launched in January 2024
Huiizon	Miao Yi Duo	Launched in April 2024
Yunsheng (a subsidiary of Hisun Animal AH)	/	Coming soon
HAPHARM GROUP	/	Coming soon
Huapai Biological Group	Mao Kang Ning	Coming soon
Keqian Biology	Ke Miao You	Coming soon

Below are some typical process routes for FVRCP vaccine, briefly listed:

The commonly used cell lines for preparing FVRCP vaccine are feline kidney cells F81 or CRFK, which belong to adherent culture cells. The cultured virus undergoes initial purification through downstream clarification filtration and ultrafiltration methods. Subsequently, chromatography is used to further reduce impurities such as HCP and HCD in the target product, thereby minimizing side effects.

The common feline distemper virus is generally above 18nm and can be effectively purified through the flow-through mode of MaXtar^® COLL400, a composite mode chromatography packing specifically designed for viruses or macromolecules. This method has flexible operating conditions, relatively easy to repeat and implement, and does not require extensive parameter exploration. Currently, a number of human and veterinary vaccines have been commercially produced using similar composite mode packing materials.

Application of MaXtar^® COLL 400 in the purification of FVRCP vaccine, (Study on the impact of different flow rates):

• Column: 16*16.7cm, CV: 33.4ml

• Buffer: 20mM PB + 0.1M NaCl, pH 6.5

Samples were loaded at flow rates of 330cm/h, 150cm/h, and 60cm/h, respectively.

Conclusion: In these three experiments, samples were loaded at flow rates of 330cm/h, 150cm/h, and 60cm/h, respectively, and the recovery rates all met expectations. MaXtar® COLL 400 can also ensure the recovery rate at low flow rates and achieved the desired results in the 100L scale-up production.

Product Introduction of MaXtar^® COLL 400/700

Featuring a unique double-layer structure design, the shell is a porous passivation layer that ensures macromolecules of 400 KD or 700 KD and above do not enter the pores and instead directly flow through the outer water path. The core is a spherical kernel coupled with groups that have both hydrophobic and negatively charged adsorbent properties, maximizing the binding of impurities such as host proteins and nucleic acids. This product boasts significant advantages:

• Processing macromolecules of 400 KD or 700 KD and above in a flow-through mode makes process optimization easier and more convenient for linear scale-up.

• The improved MaXtar^® matrix has enhanced rigidity, enabling higher process flow rates at lower backpressures, thereby improving process efficiency.

• Compared to traditional gel filtration chromatography, it offers a larger sample loading capacity, higher flow rates, and significantly enhanced production efficiency.

• It has a wide tolerance range and excellent chemical stability.

• Suitable for the separation of biological macromolecules such as viruses and viroid particles.

Tips for the Application Process of MaXtar^® COLL 400/700:

To maximize the impurity-binding effects of the various ligands within the COLL series, it is recommended to add 100~150mM NaCl to the buffer solution.

• Due to the efficient multi-mode binding between impurities and internal ligands, effective cleaning of the resin is required. It is recommended to use 1M NaOH or 30% isopropanol for this purpose.

• Citrate buffer is not recommended as citrate ions can interact with the ligands.

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