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652857 
Journal Article 
Optimisation of the coupled monoclonal antibody density for recombinant hepatitis B virus surface antigen immunopurification 
Hernandez, R; Plana, L; Gomez, L; Exposito, N; Valdes, J; Paez, R; Martinez, E; Beldarrain, A 
2005 
Yes 
Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
ISSN: 1570-0232
EISSN: 1873-376X 
816 
1-2 
1-6 
English 
15664326 
WOS:000226879800001 
Using immunosorbents based upon cyanogen bromide-Sepharose CL-4B, we have examined different ligand densities in coupling of monoclonal antibody (MAb) to find the best performance, for recombinant hepatitis B surface antigen (rHBsAg) purification. Three replicates of 5 and 15 cycles of densities ranges: 2.17-2.19, 3.18-3.62, 4.06-4.17, and 5.13-5.40 mg/ml (control); or 1.81-2.47, 3.17-3.41, 4.16-4.28, and 5.16-5.19 mg/ml (control), respectively were evaluated in terms of binding capacity, antigen recovery, ligand leakage and purity of antigen, and compared to the control. Adsorption and antigen recovery of immunosorbents manufactured were not different statistically, eventhough increased 8.08 and 9.90% at a range of 3.17-3.41 mg/ml. At this range, efficiency expressed as productivity and MAb saving was optimal. Ligand leakage and purity of antigen showed similar behaviour among all densities. Aspects related to ligand density in antigen immunoaffinity purification are discussed. 
*Antibodies, Monoclonal; Chromatography, Affinity/*methods; Cyanogen Bromide; Hepatitis B Surface Antigens/*isolation & purification; Immunosorbents; Ligands; Pichia/metabolism; Recombinant Proteins/isolation & purification; Sepharose