Recombinant Cysteine-Rich Anticoagulant (CRA) from Hirudo medicinalis — Production and C-Terminal Motif

Optimized, scalable production and purification of recombinant cysteine-rich anticoagulant protein (CRA) from Hirudo medicinalis saliva in E. coli: 3.7–5.5 mg purified protein per liter of culture across three independent experiments, with 7.1–31.2 ng bacterial proteins, 1.2–15.1 ng DNA and 60–1445 EU endotoxin per mg of protein, and strong anticoagulant activity in clotting assays. A C-terminally truncated form lost membrane affinity and had higher thrombin-time activity but lower prothrombin-time and aPTT activity than the full-length protein.

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