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Assessment of Different Regeneration and Washing Solutions in an Affinity Chromatography Procedure to Purify Plantibodies
Authors:Tatiana Alvarez  Williams Ferro  Yenisley Medina  Déborah Geada  Andrés Tamayo  Sigifredo Padilla  Mayra Wood  Tatiana González  Rodolfo Valdés
Affiliation:1. Monoclonal Antibody Production Department, Center for Genetic Engineering and Biotechnology, Ave 31/158 and 190, 10600, PO Box 6162, Havana, Cuba
2. Tobacco Research Institute, Tumbadero Road 8? km. San Antonio de los Ba?os, 3500, Havana, Cuba
3. Process Control Department, Center for Genetic Engineering and Biotechnology, Ave 31/158 and 190, 10600, PO Box 6162, Havana, Cuba
Abstract:Purification efficiency is a great challenge in plantibody production. Several regeneration?+?washing solutions of ProSep?-vA Ultra matrix used in plantibody HB-01 purification were assessed in this study to increase matrix stability without affecting purity and to reduce costs. In conclusion, PHB-01 purity was always higher than 90%. Citric acid (100?mM); pH 3.0/0.1% Triton X-100?+?150?mM PBS; pH 8.0 and 250?mM phosphoric acid; pH 1.5?+?150?mM PBS; pH 8.0/0.1% Triton X-100 allowed increasing purification cycles up to 60 and antibody yield 1.4 and 1.3 fold, respectively, together with a significant reduction in plantibody purification cost.
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