TNFRSF10B recombinant proteins refer to engineered versions of TNF Receptor Superfamily Member 10B (also known as Death Receptor 5 (DR5) or TRAIL-R2). This protein is a receptor in the TNF receptor superfamily that is involved in mediating apoptosis through interactions with its ligand, TRAIL (TNF-related apoptosis-inducing ligand). Recombinant TNFRSF10B proteins are valuable tools in studying apoptosis, cancer research, and the development of targeted therapies.

Structure and Function

• TNFRSF10B (DR5/TRAIL-R2) is a transmembrane protein with an extracellular domain that binds TRAIL, leading to receptor oligomerization and activation of apoptotic signaling pathways.
• It contains a Death Domain in its intracellular portion, which is crucial for initiating the apoptosis cascade following TRAIL binding.
• Recombinant TNFRSF10B proteins often include the extracellular domain or the full-length receptor for studying ligand binding, receptor signaling, and functional assays.

Production

• Recombinant TNFRSF10B proteins are produced in mammalian cells (such as HEK293 or CHO cells) to ensure correct folding and post-translational modifications, including glycosylation.
• Alternatively, proteins can be expressed in insect cells or yeast for certain applications.
• The gene encoding TNFRSF10B is cloned into an expression vector, and the protein is expressed and purified using affinity chromatography techniques, typically with tags like His-tag or GST-tag.

Applications

• Cancer Research: DR5/TRAIL-R2 is a target for cancer immunotherapy because of its role in inducing apoptosis in cancer cells. Recombinant TNFRSF10B proteins are used to study the interaction between TRAIL and DR5, helping to design therapies that exploit this pathway to selectively kill cancer cells.
• Drug Development: These proteins are used to screen for compounds that can enhance TRAIL-induced apoptosis or modulate DR5 signaling, aiding in the development of new cancer treatments.
• Basic Research: Recombinant TNFRSF10B proteins are crucial for understanding the molecular mechanisms of apoptosis and receptor signaling. They help in elucidating the role of DR5 in various physiological and pathological processes.
• Diagnostics: They can be used in assays to measure the levels of DR5 or to evaluate the efficacy of TRAIL-based therapies in clinical settings.

Validation and Quality Control

• Validation of recombinant TNFRSF10B proteins involves confirming their identity, purity, and functionality using methods such as SDS-PAGE, Western blotting, and mass spectrometry.
• Functional assays, including receptor-ligand binding studies and apoptosis assays, ensure that the recombinant proteins are biologically active and suitable for their intended research or therapeutic applications