Cost effective human protein C purification from Cohn Fraction IV-1 using mini-antibody.

Abstract

COST EFFECTIVE HUMAN PROTEIN C PURIFICATION FROM COHN FRACTION IV-l USING MINI-ANTIBODY Doh Gyeuhn Ahn August 5, 2005 Protein C (PC) is an important anticoagulant, antithrombotic, and antiinflammatory in blood plasma. PC deficiency can lead to severe venous thrombotic events, including lung embolism, stroke, and heart attack. In the body, PC is activated only when it is needed, and, therefore, PC does not cause bleeding problems that currently available anticoagulants may do. Purification of PC from plasma IS currently done by immunoaffinity chromatography using monoclonal antibodies (Mabs), which is very expensive. Single chain variable regions (mini-Mab) with a PC binding capability, which can b(: produced in recombinant E. coli, were developed for PC purification. Compared to Mabs, miniMabs are easier to control the contamination during the production, with lower production cost, and easier to scale-up the process. In this Ph.D. research, the effect of media conditions on the PC mini-Mab production, the mini-Mab purification yield from the production media, and the PC purification performance of the mini-Mab from Cohn Fraction IV -1 (an inexpensive PC source) were studied. The optimum conditions for mini-Mab production medium were detemlined to be 0.1 % glucose, 0.1 mM IPTG, initial pH 5.5~6.0, 23~30 °c of media temperature during