title: Comparison of different ligand densities in immunoaffinity chromatography of the plantibody HB-01 coupled to Sepharose CL-4B to purify the rHBsAg.
abstract: This paper evaluates the immunopurification behavior of a plantibody HBsAg specific plantibody coupled to Sepharose CL-4B at different ligand densities. Results show no significant differences in the adsorption and elution capacities, and rHBsAg recovery of immunosorbents at 3.43, 4.45, and 5.31 mg/mL of ligand densities compared to its mouse-derived mAb counterpart consistently used in the rHBsAg purification process. Therefore, plantibody ligand densities higher than 3.43 mg/mL do not improve the immunopurification behavior of this immunosorbent, but increase the antibody consumption and the Hepatitis B vaccine cost. Immunosorbent of 2.23 mg/mL of ligand density demonstrated a poor performance. The IgG leached detectable level never exceeded the approved limit (3 ng IgG/microg rHBsAg). Values close to this limit were only observed at the ligand density of 5.31 and 2.27 mg/mL. In the case of the ligand density of 2.23 mg/mL the IgG leached value was high (2.90 ng IgG/microg rHBsAg) due to a low level of eluted antigen. In conclusion, it supports feasibility of using this plantibody at 3.43 mg/mL of ligand density for large-scale immunopurification of rHBsAg for human use, avoiding the biosafety and ethical concerns of the massive use of animals for this purpose.
authors: Monoclonal Antibody Production Department, Center for Genetic Engineering and Biotechnology, P.O. Box 6162, Havana 10600, Cuba.
authors: Rodolfo
authors: R
authors: Valdés
authors: Yenisley
authors: Y
authors: Medina
authors: William
authors: W
authors: Ferro
authors: Biunayki
authors: B
authors: Reyes
authors: Déborah
authors: D
authors: Geada
authors: José
authors: J
authors: Montero
authors: Tatiana
authors: T
authors: Alvarez
authors: Alberto
authors: A
authors: Leyva
authors: Leonardo
authors: L
authors: Gómez
authors: Sigifredo
authors: S
authors: Padilla
authors: Leonardo
authors: L
authors: Pacín
authors: Alejandro
authors: A
authors: Figueroa
authors: Andrés
authors: A
authors: Tamayo
authors: Lorely
authors: L
authors: Milá
authors: Yurisleydi
authors: Y
authors: Aldama
authors: Galina
authors: G
authors: Moya
authors: Jorge
authors: J
authors: Reonde
authors: María del Carmen
authors: Mdel C
authors: Abrahantes
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000937:Antigen-Antibody Reactions; D015153:Blotting, Western; D002846:Chromatography, Affinity / Q000379:methods*; D002850:Chromatography, Gel / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D006514:Hepatitis B Surface Antigens / Q000302:isolation & purification*; D007124:Immunoenzyme Techniques; D008024:Ligands; D051379:Mice; D008969:Molecular Sequence Data; D046549:Plantibodies / Q000276:immunology*; D011994:Recombinant Proteins / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D012685:Sepharose / Q000737:chemistry*
chemical_list: D006514:Hepatitis B Surface Antigens; D008024:Ligands; D046549:Plantibodies; D011994:Recombinant Proteins; D012685:Sepharose