title: Simple immunoaffinity method to purify recombinant hepatitis B surface antigen secreted by transfected mammalian cells.
abstract: Purification of recombinant hepatitis B surface antigen (recHBsAg) produced in a stable Chinese hamster ovary (CHO) cell line was evaluated using Linx Affinity Purification System (Invitrogen, USA). To purify HBsAg secreted by this cell line, a murine monoclonal antibody (MAbAH1) raised against native HBsAg was used. The purified AH1MAb was conjugated with phenyldiboronic acid (PDBA) and immobilized on the immunoaffinity chromatographic support. Using an optimized protocol the affinity column was able to purify recHBsAg from supernatant of mammalian cells cultures with more than 80% purity. This method showed to be simple and quicker than the current ultracentrifugation methods. The method is also efficient and economical in obtaining purified recHBsAg.
authors.affiliation: Department of Reagents for Diagnostic, Laboratory of Hepatitis, P.O. Box 926, Bio-Manguinhos, Fiocruz, Brazil.
authors.forename: Sergio
authors.initials: S
authors.lastname: da Silva e Mouta
authors.forename: Carlos
authors.initials: C
authors.lastname: Otávio Alves Vianna
authors.forename: Ilka
authors.initials: I
authors.lastname: Ennes
authors.forename: Selma
authors.initials: S
authors.lastname: de Andrade Gomes
authors.forename: Marcos
authors.initials: M
authors.lastname: da Silva Freire
authors.forename: Edimilson
authors.initials: E
authors.lastname: Domingos da Silva
authors.forename: Salvatore
authors.initials: S
authors.lastname: Giovanni De Simone
authors.forename: Marcia
authors.initials: M
authors.lastname: Terezinha Baroni de Moraes
mesh_terms: D000818:Animals; D016466:CHO Cells; D002846:Chromatography, Affinity / Q000379:methods*; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel; D006514:Hepatitis B Surface Antigens / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000302:isolation & purification; D014162:Transfection
chemical_list: D006514:Hepatitis B Surface Antigens; D011994:Recombinant Proteins