Comparison of different elution conditions for the immunopurification of recombinant hepatitis B surface antigen.
An immunoaffinity chromatographic method was developed using a mAb immunosorbent to purify recombinant hepatitis B surface antigen (r-HBsAg) from yeast. Elution conditions using a mAb-coated ELISA were improved to select the best conditions to purify r-HBsAg. The optimum results in terms of total quantitative recovery were obtained using 20 mM Tris pH 11.6. An increase in the CB.Hep-1 mAb (anti-HBsAg) useful immunosorbents half-life and in its yield per cycle was obtained when alkaline elution conditions were used. Moreover, the basic conditions do not affect either the antigenic characteristics or the purity or the molecular integrity of r-HBsAg.
Ibarra
N
N
Monoclonal Antibodies Division, Center for Genetic Engineering and Biotechnology, Havana, Cuba.
Caballero
A
A
González
E
E
Valdés
R
R
D002846:Chromatography, Affinity / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D006514:Hepatitis B Surface Antigens / Q000302:isolation & purification*; D007202:Indicators and Reagents; D011994:Recombinant Proteins / Q000302:isolation & purification
D006514:Hepatitis B Surface Antigens; D007202:Indicators and Reagents; D011994:Recombinant Proteins