Abstract
Antigen Used for Immunization
Recombinant 6XHis-tagged maltose binding protein expressed in Escherichia coli using plasmid vector pDB.His.MBP (DNASU).
Method of Immunization
Two hundred microliters prepared solution (50 μg of purified recombinant 6XHis-MBP supplemented with 50 μg poly [I:C]) and 50 μg of antimouse CD40 monoclonal antibody as adjuvant were injected intraperitoneally into female BALB/c mice (8 weeks of age). The mice were injected twice with a 2-week interval and sera were tested by Western blot. Three days before fusion, the final boost was injected through the tail vein.
Parental Cell Line Used for Fusion
Mouse myeloma cell line Sp2/0-Ag14.
Selection and Cloning Procedure
Selection by HAT (hypoxanthine–aminopterin–thymidine) media, cloned by flow cytometry and positive clone identification by dot blot.
Heavy and Light Chains of Immunoglobulin
Heavy chain, gamma1; light chain, kappa.
Specificity
Validation and specificity were determined by dot blot, Western blot, immunoprecipitation, and ELISA.
Specific Antigen Identified
E. coli maltose binding protein.
Availability
Monoclonal antibodies DSHB-MBP-3D7 and DSHB-MBP-2A1 are available from the Developmental Studies Hybridoma Bank.