Development of a bispecific monoclonal antibody as a universal immunoprobe for detecting biotinylated macromolecules.
Bispecific monoclonal antibody (BsMab) combining two different antigen binding sites, anti-biotin and anti-HRPO paratopes, could be used as a universal immunoprobe for detecting all biotinylated macromolecules. First, a mouse hybridoma cell line secreting monospecific anti-biotin Mab was generated and characterized. Second, a quadroma cell line which could continuously secrete bsMab (anti-biotin x anti-HRPO) was developed by a nonselective microelectrofusion method. The supernatant containing bsMab was collected from tissue culture medium and purified with two affinity columns. This bsMab has comparable avidity to commercial streptavidin-HRPO when tested against biotinylated macromolecules. Compared to streptavidin, this bsMab can bind the enzyme and thus eliminate the need for chemical conjugation. This bsMab can be used as a promising immunoprobe for detecting many macromolecules bearing biotin markers, such as protein, phage, liposome and DNA in different bioassay systems.
Cao
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Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Edmonton, Canada.
Christian
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Suresh
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D000818:Animals; D018033:Antibodies, Bispecific / Q000276:immunology* / Q000302:isolation & purification; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D000915:Antibody Affinity; D001666:Binding Sites, Antibody; D001710:Biotin / Q000276:immunology*; D019921:Biotinylation; D015153:Blotting, Western; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification*; D006735:Horseradish Peroxidase / Q000276:immunology*; D006825:Hybridomas / Q000276:immunology; D007118:Immunoassay / Q000379:methods*; D008081:Liposomes; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*; D051381:Rats
D018033:Antibodies, Bispecific; D000911:Antibodies, Monoclonal; D008081:Liposomes; D011506:Proteins; D001710:Biotin; D004247:DNA; D006735:Horseradish Peroxidase