Erythropoietin (EPO), known as red blood cell stimulating factor, has the main biological functions of inhibiting myelocyte apoptosis, stimulating the release of reticulocytes into the blood circulation, and stimulating protein synthesis within cells. It is the main hematopoietic growth factor responsible for the proliferation, differentiation, and maintenance of normal peripheral circulating red blood cells in the body. Recombinant human erythropoietin (rhEPO) and natural erythropoietin (EPO) have very similar structures, with only slight differences in sialic acid linkage. In clinical medicine, rhEPO is mainly used to treat anemia caused by renal insufficiency, anemia of AIDS patients, anemia related to malignant tumors and other diseases.

rhEPO Process

At present, we mainly use genetic engineering technology to recombine the EPO gene in vitro and introduce it into CHO cells. The CHO cells are cultured for expression, and then purified through multiple steps to obtain recombinant human erythropoietin (rhEPO). The use of CHO cells as the production vector for rhEPO has a potential risk of viral contamination. How to robustly control the potential risk of viral contamination is of utmost importance in the production process of rhEPO.

According to relevant regulations and guidelines domestic and overseas, the design and implementation of virus clearance research in the same production process require the use of different virus inactivation or removal methods to ensure that biological products do not carry any potential risk of virus contamination when used for treating or preventing diseases. Chromatography and virus removal filtration can both be effective virus removal steps, among which virus removal filtration doesn’t change the original physical and chemical properties of the material, and has a high LRV removal effect, making it an ideal method for virus contamination control.

EPO Process Filtration by Cobetter Virus Removal Filters

Case Study 1

Condition

• Filterability test of 1.5 g/L EPO
• Prefiltration by PDT and PNY prefilters, respectively
• Pre-filtered samples are tested by Cobetter Viruclear VF Plus at 30psi

Results

→PDT+VF Plus, recovery rate 97.7%
→PNY+VF Plus, recovery rate 99.2%

Case Study 2

Condition

• Filterability test of 4 g/L EPO
• Prefiltration by PDT and PNY prefilters, respectively
• Pre-filtered samples are tested by Cobetter Viruclear VF Plus at 30psi

Results

→PDT+VF Plus, recovery rate 97.3%
→PNY+VF Plus, recovery rate 99.0%

Case Study 3

Filterability test of 8 g/L EPO

Two-group Tests

• Prefiltration by PDT. The pre-filtered samples are tested by Cobetter Viruclear VF Plus at 30psi
• Prefiltration by PNY. The pre-filtered samples are tested by Cobetter Viruclear RC H at 29psi

Results
→PDT+VF Plus, recovery rate 97.9%
→PNY+RC H, recovery rate 99.8%

For the virus removal filtration process of rhEPO, the VF Plus is recommended for low-concentration, low-fouling EPO materials. For materials with higher protein concentration and high fouling potential, the Plus X or RC H virus removal filters are recommended.

For filtration processes with low flux decay, the PNY nylon prefilter is recommended. For processes experiencing significant flux decay, the depth prefilter PDT is suggested. It is important to note that depth prefilter may have higher adsorption potential, and product yield should be carefully evaluated during the process development stage.

Virus safety, as a top priority in the production process of the biopharmaceutical industry, also requires comprehensive evaluation of its removal process. Cobetter has launched three PES (polyethersulfone) virus removal filtration products and two RC (regenerated cellulose) virus removal filtration products, equipped with virus removal prefilters, to ensure the safety of biological products against virus.

In the future, we will continuously upgrade and optimize the manufacturing process of membrane filtration components to improve filtration performance and stability. We are dedicated to providing filtration solutions with high loading capacity and low cost to our customers.