Separation Of Protein C From Cohn Fraction Iv-1 By Mini-Antibody
Human protein C (PC) is a natural anticoagulant, antithrombotic, anti-inflammatory, and anti-apoptotic in the bloodstream. PC deficiency can lead to abnormal blood clot formation inside blood vessels, possibly causing heart attack, stroke, skin necrosis, or even death. PC can be, therefore, a valuable therapeutic with little side effect, unlike the currently used anti-coagulants. To reduce the cost involved in immuno purification of PC from blood plasma, single chain variable fragments (mini-Mab) are being produced by recombinantE. coli using phagemid technique. As an economic means of purifying the PC specific mini-Mab, metal affinity chromatography (IMAC) purification process was also investigated. Then using the purified mini-Mab, the feasibility of PC purification from the Cohn Fraction IV-1 was examined. Cohn Fraction IV-1 is usually a discarded side-stream from the blood plasma fractionation of human serum albumin. It holds 90% of PC in plasma, but is very cheap. Preliminary study of PC purification from the Cohn Fraction IV-1 showed 16% purification yield using mini-Mab immobilized NHS-activated Sepharose. The economic analysis for PC purification using mini-Mab showed that the overall process was found to be tens of times cheaper than that using Mab.
KeywordsHuman Serum Albumin Elution Step Single Chain Variable Fragment Purification Yield Urea Hydrogen Peroxide
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- 2.L. K. Korah and K. A. Kang, Prelimanary Study for the Protein C Purification Using Mini-Antibodies Produced from Recombinant E. coli, Oxygen Transport to Tissue XXV.Thorniley, Harrison, and James (Kluwer Academic/Plenum Publishers, 2003), pp. 171-176.Google Scholar
- 4.P. Verheesen, C. T. Verrips, and J. W. Haard, Beneficial Properties of Single-domain Antibody Fragments for Application in Immuno affinity Purification and Immuno-perfusion Chromatography, Biochimicaet Biophysica. Acta. 1624, 21-28 (2003).Google Scholar
- 6.S. Rezania, D. G. Ahn, K. A. Kang, Cost Effective Metal Affinity Chromatography for Protein C Specific, Mini-Antibody Purification, Proceedings of the Annual 2004 ISOTT meeting [In press].Google Scholar
- 7.A.N.H. Creager, ‘What Blood Told Dr Cohn’: World War II, Plasma Fractionation, and the Growth of Human Blood Research, Stud. Hist. Phil. Biol. Biomed. Sci. 30, 377-405.Google Scholar