Expression, purification, and characterization of active recombinant prostate‐specific antigen in Pichia pastoris (yeast)

Prostate‐specific antigen (PSA), a member of the kallikrein family of serine proteases, is a chymotrypsin‐like glycoprotein produced by the prostate epithelium. Elevated serum PSA (> 4 ng/ml) is a tumor marker for prostatic cancer and benign prostatic hypertrophy; increasing serum PSA over time is indicative of metastatic disease. It has been suggested that PSA may contribute to tumor metastasis through degradation of extracellular matrix glycoproteins, as well as cleavage of IGF binding protein‐3, a modulator of IGF‐1. To elucidate the role of PSA in the development and progression of prostatic cancer, it is necessary to have a reliable, cost‐effective source of enzymatically active protein. Previous efforts to express recombinant PSA (rPSA) produced inactive proPSA, or mixtures of active and inactive PSA requiring activation by removal of the propeptide. We describe the expression of active recombinant mature PSA in yeast.

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