부산대학교 도서관

Plantibody purification is not as e fficient as antibody purification from serum, ascites, or mammalian cell cultures. It is characterized by the application of inefficient plantibody solid-liquid extraction systems, low plantibody recovery, and short lifetimes of expensive chromatography matrices. To overcome it, several protocols of liquid-liquid aqueous two-phase extraction (ATPE) combined with a ffinity chromatography were previously studied to purify the CB.Hep-1 monoclonal antibody, which showed an unexpectedly high recovery. However, a study of ATPE combined with several a ffin ity chromatography matrices to purify plantibodies has not been reported so far. Therefore, a combination o f the best ATPE protocol w ith five specific a ffin ity chromatography matrices to purify a plantibody for vaccine manufacturing is described in this study. Positive outcomes from plantibody recovery (%), specific activity (%), yield (mg purified IgG/L of leaf extract), and productivity (mg purified IgG/L of leaf extract/h) were achieved. Plantibody purity did not show statistical differences among all samples (>97%, p<0.05), and protein A leakage was thousands of times smaller than toxic protein A for nonhuman primates. In summary, the combination of ATPE (10% PEG 4000/15% K2P04, pH 5.5) with two specific affinity resins were well-suited for large-scale plantibody purification from tobacco plant leaves. [ABSTRACT FROM AUTHOR]