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 recombinant E. 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.
Rezania
Samin
S
Department of Chemical Engineering, University of Louisville, Louisville, KY 40292, USA.
Ahn
Doh G
DG
Kang
Kyung A
KA
D000911:Antibodies, Monoclonal / Q000378:metabolism*; D001798:Blood Proteins / Q000737:chemistry / Q000378:metabolism; D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods; D006801:Humans; D011486:Protein C / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal; D001798:Blood Proteins; C032309:Cohn fraction IV; D011486:Protein C