Assessment of Affinity Chromatography Matrices in Plantibody Purification from Aqueous Two-Phase Extraction Samples

by Williams Ferro, Tatiana Alvarez, Déborah Geada, Yenisley Medina, José Montero, Andrés Tamayo, Ariadna López, Daily Hernández, Mayra Wood, Tatiana González, Regla Somoza, José García, Dobián Cecilia, Yanara González, Osmaro González, David Gavilán, Mailyn LaO, and Rodolfo Valdés
Volume 15, Issue 4 (Winter 2016/2017)

Plantibody purification is not as efficient 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 affinity 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 affinity chromatography matrices to purify plantibodies has not been reported so far. Therefore, a combination of the best ATPE protocol with five specific affinity 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 non-human primates. In summary, the combination of ATPE (10% PEG 4000/15% K2PO4, pH 5.5) with two specific affinity resins were well-suited for large-scale plantibody purification from tobacco plant leaves...

Ferro W, Alvarez T, Geada D, Medina Y, Montero J, Tamayo A et al. Assessment of affinity chromatography matrices in plantibody purification from aqueous two-phase extraction samples. BioProcess J, 2017; 15(4): 43–51.

Posted online February 15, 2017.