Procurement of reference materials against recombinant HBsAg surface antigen.
Mabs against HBsAg have been used for structural analyses, development of diagnostic tests, and for antigen immunopurification. Resultant products obtained from current methods of genetic recombination demand reference materials to test their potency, identity, purity, and the biological and immunological specific activity corresponding to their manufacturing processes. In this paper, we present a method for the qualitative and quantitative characterisation of CB.Hep-1 and CB.Hep-4 RM and their stability, in real time, with the required quality to be used as primary reference materials. Among the criteria applied in this study, we considered Mab-specific concentration through ELISA, purity, and total proteins by various methods, quantification of antigen recognition capacity of Mabs through the Ag-Mab absolute-recognition method. Sterility, homogeneity, stability, subclass, and isoelectric focusing were used in the characterisation of the reference materials.
Abrahantes
Maria del Carmen
Mdel C
Process Control Division, Centre for Genetic Engineering and Biotechnology, Havana, Cuba.
Reyes
Biunayki
B
Reyes
Jesus
J
Sosa
Raudel
R
González
Ernesto M
EM
Vega
Maribel
M
Valdés
Rodolfo
R
Martínez
Eduardo
E
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification; D001202:Ascitic Fluid / Q000276:immunology; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006510:Hepatitis B Antibodies / Q000096:biosynthesis / Q000276:immunology; D006514:Hepatitis B Surface Antigens / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D007074:Immunoglobulin G / Q000276:immunology*; D007132:Immunoglobulin Isotypes / Q000032:analysis / Q000276:immunology; D007525:Isoelectric Focusing; D016014:Linear Models; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification; D012015:Reference Standards; D012756:Sheep
D000911:Antibodies, Monoclonal; D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens; D007074:Immunoglobulin G; D007132:Immunoglobulin Isotypes; D011994:Recombinant Proteins