Comparison of different ligand densities in immunoaffinity chromatography of the plantibody HB-01 coupled to Sepharose CL-4B to purify the rHBsAg.
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.
Valdés
Rodolfo
R
Monoclonal Antibody Production Department, Center for Genetic Engineering and Biotechnology, P.O. Box 6162, Havana 10600, Cuba.
Medina
Yenisley
Y
Ferro
William
W
Reyes
Biunayki
B
Geada
Déborah
D
Montero
José
J
Alvarez
Tatiana
T
Leyva
Alberto
A
Gómez
Leonardo
L
Padilla
Sigifredo
S
Pacín
Leonardo
L
Figueroa
Alejandro
A
Tamayo
Andrés
A
Milá
Lorely
L
Aldama
Yurisleydi
Y
Moya
Galina
G
Reonde
Jorge
J
Abrahantes
María del Carmen
Mdel C
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*
D006514:Hepatitis B Surface Antigens; D008024:Ligands; D046549:Plantibodies; D011994:Recombinant Proteins; D012685:Sepharose