Stirrer tank: an appropriate technology to immobilize the CB.Hep-1 monoclonal antibody for immunoaffinity purification.
The CB.Hep-1 monoclonal antibody was coupled to CNBr-activated Sepharose CL 4B at three different immobilization scales for purification of recombinant hepatitis B surface antigen. Standard laboratory apparatus to obtain immunosorbents of 1 l (scale I) and 3 l (scale II) as well as a stirrer tank to prepare 6 l immunosorbents (scale III) were used. The binding capacity at scale III was 2- and 1.5-fold higher with respect to the scales II and I, while a reduction in the ligand leakage of 5- and 2-folds was observed. Immunosorbents from scale II showed a significantly reduced adsorption, and an increased ligand leakage. Differences in the coupling efficiency were not observed. Antigen purity eluted from the immunosorbents was always above 85%.
Hernández
R
R
Monoclonal Antibody Division, Center for Genetic Engineering and Biotechnology, Havana, Cuba. raul.hernandez@cigb.edu.cu
Chong
E
E
Morales
R
R
Pérez
E
E
Amador
Y
Y
Zubiaurrez
J R
JR
Valdés
R
R
Figueroa
A
A
Agraz
A
A
Herrera
L
L
D000911:Antibodies, Monoclonal; D006514:Hepatitis B Surface Antigens / Q000276:immunology / Q000302:isolation & purification*; D007163:Immunosorbent Techniques / Q000295:instrumentation*
D000911:Antibodies, Monoclonal; D006514:Hepatitis B Surface Antigens