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

Mode of Action of tPA:
Tissue Plasminogen Activator (tPA) is an enzyme that catalyzes the conversion of plasminogen, an inactive plasma protein, into plasmin, an active enzyme responsible for breaking down fibrin clots (fibrinolysis). This property makes tPA useful in treating blood clots during heart attacks and strokes by dissolving clots and restoring normal blood flow. Production of tPA through Mammalian Cell Culture: tPA is produced using recombinant DNA technology in mammalian cells (e.g., Chinese Hamster Ovary (CHO) cells), which can perform proper post-translational modifications necessary for its activity. Schematic representation:

1. Isolation of the human tPA gene.
2. Insertion of tPA gene into a suitable expression vector.
3. Introduction of recombinant vector into mammalian host cells (e.g., CHO cells) by transfection.
4. Culturing transfected cells under suitable conditions to express tPA protein.
5. Harvesting and purification of tPA protein from culture medium.

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