SERPINC1 (Serpin Family C Member 1), also known as Antithrombin III (ATIII), is a crucial plasma protein that inhibits several proteases in the blood clotting cascade, particularly thrombin and factor Xa. It plays a vital role in regulating blood coagulation and preventing excessive clot formation. Deficiency or dysfunction of SERPINC1 can lead to thrombotic disorders or bleeding complications.

Recombinant SERPINC1 Protein

Recombinant SERPINC1 protein is produced using recombinant DNA technology. This involves inserting the SERPINC1 gene into an expression vector, transforming it into a suitable host cell system, and expressing and purifying the protein.

Production Process

• Gene Cloning: The SERPINC1 gene is cloned into an expression vector, which may include additional sequences for protein purification and stabilization.
• Transformation: The vector is introduced into a host cell system (e.g., E. coli, yeast, or mammalian cells) using techniques such as heat shock, electroporation, or transfection.
• Expression: The host cells are cultured under conditions that induce the expression of the recombinant SERPINC1 protein.
• Purification: The protein is purified from the cell lysate or culture supernatant using chromatography techniques, such as affinity chromatography or ion exchange chromatography, to achieve high purity and functionality.

Structure and Function

• Structure: SERPINC1 is a serine protease inhibitor (serpin) with a single-chain structure that includes a reactive center loop. The protein undergoes a conformational change upon binding to its target proteases, which leads to their inhibition.
• Function: SERPINC1 inhibits several key enzymes in the coagulation cascade, including thrombin and factor Xa, by forming a stable complex with them. This inhibition is essential for maintaining the balance between clot formation and dissolution, thus preventing excessive clotting and ensuring normal blood flow.

Applications

• Research: Used to study the mechanisms of blood coagulation and the role of SERPINC1 in regulating protease activity. It also helps investigate the effects of SERPINC1 deficiencies or mutations.
• Therapeutics: Recombinant SERPINC1 can be used as a therapeutic agent to treat conditions associated with SERPINC1 deficiency, such as thrombosis or bleeding disorders. It can also be used to develop and test new treatments for coagulation disorders.
• Diagnostics: Employed in assays to measure SERPINC1 levels or activity in clinical samples, aiding in the diagnosis and management of coagulation disorders.

Quality Control

• Purity: Verified by SDS-PAGE, Western blotting, and HPLC to ensure the absence of contaminants.
• Activity: Confirmed through functional assays, such as thrombin inhibition assays, to ensure the protein retains its ability to inhibit its target proteases effectively.
• Stability: Assessed under various storage conditions to ensure the protein maintains its functional integrity over time.

Recombinant SERPINC1 protein is a critical tool for studying and managing coagulation disorders. Its production and application in research, therapeutic development, and diagnostics contribute significantly to understanding and treating conditions related to blood clotting and antithrombin deficiencies.