TFF Technology Applicates in Purification Of Cytokine-based Drugs
Cytokine therapy, as a cutting-edge technology in current medical field, is gradually showing its powerful therapeutic potential. Cytokines are produced by immune cells and related cells. This treatment mainly uses cytokines to fight disease directly or indirectly, especially cancer, by regulating cell function.
In clinical practice, cytokine therapy has shown efficacy in a variety of diseases. In addition to cancer treatment, it is also used in autoimmune diseases, viral infections and other fields. In addition, cytokines also play an important role in detection, cosmetic medicine and other fields.
The currently approved cytokine-based drugs include IFN-α, IFN-β, IFN-γ, IFN-ε, PO, GM-CSF, G-CSF, IL-3; The cytokine-based drugs in clinical trials, include IL-1, IL-3, IL-4, IL-6, IL-11, M-CSF, SCF, TGF-β, etc. They increase cytokines in the body through various ways to achieve the purpose of treating diseases. IFN, IL-2, TNF, CSF are better in clinical treatment.
IFN (interferon) is a kind of low molecular weight proteins (mainly glycoproteins) with a variety of biological activities. It does not kill or inhibit viruses directly, but inhibits viral replication by inducing cell surface receptors and making cells produce antiviral proteins. It is the most important cytokine for antiviral infection. According to the different structure, receptor and function, IFN is mainly divided into type I, type II, type III, and non-classical type IV (IFN-λ4) has also been found in recent years. Type I interferon members include IFN-α (13 subtypes), IFN-β, IFN-ε, IFN-κ, IFN-ω, etc. Type II interferon members are only IFN-γ. Type III interferon members are IFN-λ1 (IL-29), IFN-λ2 (IL-28A), IFN-λ3 (IL-28B), and IFN-λ4 (unique to humans).
IL-2 (interleukin-2) is a pleiotropic cytokine produced by activated T cells (primarily CD4+ Th1 cells). It plays a central role in immune regulation, T cell proliferation, anti-tumor immunity, and autoimmune tolerance. IL-2 has been widely used in clinic, especially in the treatment of tumor, such as kidney cancer and melanoma. It also plays an important role in autoimmune diseases treatments, such as type 1 diabetes, lupus erythematosus and other autoimmune diseases.
TNF (tumor necrosis factor) is a kind of pleiotropic cytokines primarily produced by immune cells. TNF plays a complex role in inflammation, immune regulation, apoptosis and tumorigenesis. It is not only a protective factor, but also may participate in pathological processes. Its core biological functions include active inflammatory response, regulation of cell survival and death, and mediating immune homeostasis. There are two main types of TNF, including TNF-α and TNF-β.
CSF (colony stimulating factor) is a kind of cytokine that regulates and controls the proliferation, differentiation and immune cell function of hematopoietic stem cells. According to different target cells, it is mainly divided into G-CSF, GM-CSF (granulocytic-macrophage colony stimulating factor) and M-CSF, which play a key role in blood system homeostasis, infection defense and tumor therapy.
The downstream purification of cytokine-based drugs is a highly complex and precise process, and its pathways are different because of the difference of expression systems. It is worth noting that tangential flow filtration (TFF), as a key generic technology, plays an irreplaceable core role in the downstream purification process of various expression systems.
The downstream purification of cytokine-based drugs typically includes several key steps: fermentation culture and harvest, microfiltration clarification (TFF, remove large particles of impurities, prepare for subsequent purification steps), ultrafiltration (TFF, reduce chromatographic processing volume, improve chromatographic efficiency, protect chromatographic columns), capture and preliminary purification, fine purification, ultrafiltration (TFF, remove small molecular impurities and replace buffers), bactericidal filtration and preparations.
Prokaryotic expression system (E. coli) has the advantages of low cost, short culture cycle and high fermentation density, which is suitable for large-scale industrial production. The genetic background is clear and the carrier system is mature, which is convenient for gene manipulation and expression optimization. Protein production can reach 10-30% of total protein; Downstream purification is relatively simple, especially the inclusion form is easy to enrich and purify. However, it should be noted that it lacks eukaryotic post-translational modification, some proteins need to be renaturated, and endotoxin should be strictly removed. It is suitable for the production of cytokines with simple structure and no need for glycosylation.
Compared with prokaryotic expression of cytokines, the formation and renaturation of inclusion bodies are avoided during the expression of mammalian cells, and the natural correct folding conformation and post-translational modification are beneficial to increase specific activity.
TFF is a dynamic filtration technology, the core feature is that the feed liquid flows tangentially along the membrane surface, rather than vertically. This flow mode can reduce the membrane surface concentration polarization and pollution effectively, thereby improve the filtration efficiency and the service life of the membrane.
Compared to other filtration technologies, TFF has the following significant advantages in the purification of cytokine-based drugs:
1. High efficiency: TFF can process a large amount of liquid quickly, shorten the purification time significantly.
2. High recovery rate: TFF can achieve higher product recovery rate due to reduced membrane contamination and concentration polarization.
3. Flexibility: TFF system can adjust the operating parameters (such as tangential flow rate, transmembrane pressure) according to different purification needs to adapt to different production scales.
4. Scalability: TFF technology is easy to scale up from the laboratory scale to the industrial production scale to meet the needs of commercial production.
In practical applications, in order to guarantee the efficiency and stability of TFF technology, it is necessary to focus on the following points:
1. Selection of membrane materials: the material of TFF membrane (such as polyether sulfone, regenerated cellulose) and the molecular weight cut-off (MWCO) have an important impact on the purification. The selection of suitable membrane materials and pore sizes is a prerequisite to efficient purification.
2. Optimization of operating parameters: tangential flow rate, transmembrane pressure, feed liquid concentration and other operating parameters need to be optimized according to specific process. Too high pressure may lead to membrane contamination, while too low flow rate may reduce filtration efficiency.
3. Cleaning and maintenance of the system: Regular cleaning and maintenance of the TFF system can extend the service life of the membrane and guarantee the stability and repeatability of the purification process.
4. Process validation and quality control: In the production of cytokine-based drugs, TFF process needs to be rigorously verified to ensure that it complies with Good Manufacture Practice (GMP).
With the rapid development of the biopharmaceutical industry, the demand for cytokine-based drugs is increasing, and the requirements for downstream purification technologies are also raising. Tangential flow filtration technology will continue to play an important role in the purification of cytogenetic drugs due to its high efficiency, flexibility and scalability.
In the future, with the continuous improvement of membrane materials and TFF systems, tangential flow filtration technology will further improve purification efficiency, reduce production costs, and provide technical support for the advent of more efficient and safe cell factor drugs.
If you have more questions about cytogenic drug purification or tangential flow filtration technology, please leave a comment! Don't forget to like, forward, let more people know about this important biopharmaceutical technology!
About Guidling
Guidling Technology is a production-oriented and high-tech enterprise focusing on the downstream clarification, separation and purification of biopharmaceuticals. The products are widely used in the filtration process of MAB, vaccine, diagnosis, blood products, serum, endotoxin and other biological products; Guidling Technology has "cassettes filter and tangential flow filtration device", "hollow fiber membrane", "virus filter", "deep membrane", "sterilizaation filter", "centrifugal filter devices" and other products, and has a large number of product lines, from small disposable laboratory filtration to production filtration system, meet the needs of testing and production. Guidling Technology is looking forward to cooperating with you!
