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/...

Full description

Saved in:
Bibliographic Details
Published in:Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2007-06, Vol.852 (1), p.1-7
Main Authors: Valdés, Rodolfo, Medina, Yenisley, Ferro, William, Reyes, Biunayki, Geada, Déborah, Montero, José, Alvarez, Tatiana, Leyva, Alberto, Gómez, Leonardo, Padilla, Sigifredo, Pacín, Leonardo, Figueroa, Alejandro, Tamayo, Andrés, Milá, Lorely, Aldama, Yurisleydi, Moya, Galina, Reonde, Jorge, Abrahantes, María del Carmen
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Analysis
Analytical, structural and metabolic biochemistry
Animals
Antigen-Antibody Reactions
Biological and medical sciences
Blotting, Western
Chromatography, Affinity - methods
Chromatography, Gel - methods
Electrophoresis, Polyacrylamide Gel
Fundamental and applied biological sciences. Psychology
General pharmacology
Hepatitis B Surface Antigens - isolation & purification
Hepatitis B virus surface antigen
Immunoaffinity
Immunoenzyme Techniques
Immunopurification
Ligands
Medical sciences
Mice
Molecular Sequence Data
Monoclonal antibody
Pharmacology. Drug treatments
Plant-derived antibody
Plantibodies - immunology
Plantibody
Recombinant Proteins - isolation & purification
Sensitivity and Specificity
Sepharose - chemistry
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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/μg 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/μg 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.
ISSN:1570-0232
1873-376X
DOI:10.1016/j.jchromb.2006.12.018