Optimisation of the coupled monoclonal antibody density for recombinant hepatitis B virus surface antigen immunopurification.
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.
Hernández
Raúl
R
Department of Purification Development Center for Genetic Engineering and Biotechnology, P.O. Box 6162, Havana 10600, Cuba. letypml@iift.cu
Plana
Liuba
L
Gómez
Leonardo
L
Expósito
Néstor
N
Valdés
Jorge
J
Páez
Rolando
R
Martínez
Eduardo
E
Beldarraín
Alejandro
A
D000911:Antibodies, Monoclonal; D002846:Chromatography, Affinity / Q000379:methods*; D003488:Cyanogen Bromide; D006514:Hepatitis B Surface Antigens / Q000302:isolation & purification*; D007164:Immunosorbents; D008024:Ligands; D010843:Pichia / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification; D012685:Sepharose
D000911:Antibodies, Monoclonal; D006514:Hepatitis B Surface Antigens; D007164:Immunosorbents; D008024:Ligands; D011994:Recombinant Proteins; D012685:Sepharose; D003488:Cyanogen Bromide