TNF recombinant proteins refer to engineered versions of Tumor Necrosis Factor (TNF), a pro-inflammatory cytokine that plays a critical role in regulating immune responses, inflammation, and cell death. TNF is involved in a range of physiological processes and diseases, including autoimmune disorders, cancer, and infectious diseases. Recombinant TNF proteins are used extensively in research, diagnostics, and therapeutic development.

Structure and Function

• TNF exists mainly in two forms: TNF-alpha (TNF-α) and TNF-beta (TNF-β). TNF-α is the most studied form and is involved in systemic inflammation, while TNF-β (also known as Lymphotoxin-alpha) is more involved in lymphoid organ development and immune regulation.
• TNF is a homotrimeric protein that binds to its receptors, TNFR1 and TNFR2, leading to various downstream effects, including apoptosis, inflammation, and modulation of immune responses.
• Recombinant TNF proteins usually include the full-length cytokine or specific domains that are crucial for receptor binding and activity.

Production

• TNF recombinant proteins are typically produced in E. coli, yeast, insect cells, or mammalian cells, depending on the need for post-translational modifications such as glycosylation and correct protein folding.
• The gene encoding TNF is cloned into an expression vector, followed by protein expression and purification using affinity chromatography techniques, often utilizing tags like His-tag or GST-tag to facilitate purification.

Applications

• Research: Recombinant TNF proteins are used to study TNF’s role in inflammation, immune responses, and cell death. They help in understanding the mechanisms of diseases where TNF is a key player, such as rheumatoid arthritis, Crohn’s disease, and cancer.
• Therapeutic Development: TNF is a target for therapeutic interventions. Recombinant TNF proteins are used to develop and test anti-TNF drugs (e.g., infliximab, adalimumab) designed to inhibit TNF activity and treat inflammatory conditions.
• Diagnostics: Recombinant TNF proteins are used in assays to detect and quantify TNF levels in patient samples, providing valuable information for diagnosing and monitoring diseases associated with abnormal TNF activity.
• Vaccine Research: They are used in vaccine development to study the immune response and to assess how TNF can modulate vaccine efficacy.

Validation and Quality Control

• Validation of recombinant TNF proteins includes confirming their identity, purity, and biological activity using techniques such as SDS-PAGE, Western blotting, and mass spectrometry.
• Functional assays, such as receptor-binding studies or bioassays that measure cytokine activity (e.g., cell proliferation or apoptosis assays), ensure that the recombinant proteins are biologically active and suitable for their intended applications.