Question

What is the mode of action of tissue Plasminogen Activator (tPA)? Draw a schematic representation to show the method of production of tPA through mammalian cell culture.

Hint:

tPA converts plasminogen to plasmin to dissolve blood clots; produced via recombinant DNA technology in mammalian cell cultures for correct folding and modification.

Answer 1

tPA Mechanism:
Tissue Plasminogen Activator (tPA) is an enzyme that converts inactive plasminogen to active plasmin. Plasmin breaks down fibrin clots through fibrinolysis. This action is utilized in treating heart attacks and strokes by dissolving clots and re-establishing blood flow. tPA Production via Mammalian Cell Culture: Recombinant DNA technology is employed to produce tPA in mammalian cells, such as Chinese Hamster Ovary (CHO) cells. These cells perform essential post-translational modifications required for tPA activity. Production Steps:

1. Obtain the human tPA gene.
2. Insert the tPA gene into an expression vector.
3. Transfect mammalian host cells (e.g., CHO cells) with the recombinant vector.
4. Culture transfected cells to express tPA protein.
5. Harvest and purify tPA from the culture medium.

% You can insert a simple diagram here if needed using TikZ or includegraphics commands.