Protein C is an important
serine protease due to its ability to proteolytically cleave activated Factors V and VIII. Excess coagulation and blood agglutination can lead to plugged capillaries, thereby reducing
oxygen transport to interstitial tissues. To treat patients with hereditary and
acquired protein C deficiency would require a greater amount of
Protein C than that available from human plasma. However, the potential demand for this
protein could be met by the production of human
protein C from transgenic animal mammary glands. Thus, research into inexpensive, efficient methods to purify
proteins from transgenic animal milk will be a critical area of study for the large scale production of
protein C. Immobilized
metal affinity chromatography (
IMAC) is a novel method for the purification of
protein C. A proposed method of purification is to take advantage of
protein C's strong
metal ion binding characteristics with
IMAC to assist in the separation from transgenic animal milk. The separation procedure is benchmarked against current systems in use by the American Red Cross for purification of
Protein C from transgenic porcine milk. Common problems in developing separation schemes for new
therapeutics are the initial availability of the product (
protein), and time-to-market concerns. Extensive experimental tests for scaleable purification schemes are often cost and time prohibitive. In order to optimize an
IMAC protocol with minimal waste of time and resources, total quality management tools have been adopted. Initial experiments were designed to choose
buffer conditions, eluents, immobilized valence metals, and flow rates using Taguchi experimental design, which is a total quality management (TQM) tool. One of the values of Taguchi methods lies in the use of Latin orthogonal sets. Through the use of the orthogonal sets, the total number of experiments may be reduced, shortening the focus time on optimal conditions.