Assay of the polyamine spermine by a monoclonal antibody-based ELISA.
Spermine-specific monoclonal antibodies were prepared by immunising mice with protein-spermine conjugates. A resulting monoclonal antibody, IAG-1, exhibited both high affinity for spermine (binding constant 5.5 x 10(7) M-1) and high specificity, cross-reacting only weakly with spermidine. Using this antibody a competitive ELISA was developed with a detection limit of 1 pmol. The assay has been used to quantify spermine content of plant tissues, without derivatization, and producing within 6 h of collection, values for the spermine content which are similar to published data obtained by HPLC.
Garthwaite
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Department of Biochemistry, University of London, Egham, Surrey, UK.
Stead
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Rider
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D000818:Animals; D000911:Antibodies, Monoclonal; D000915:Antibody Affinity; D000918:Antibody Specificity; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006825:Hybridomas; D051379:Mice; D008807:Mice, Inbred BALB C; D010047:Ovalbumin; D012709:Serum Albumin; D013095:Spermidine / Q000276:immunology; D013096:Spermine / Q000032:analysis* / Q000276:immunology; D014908:Triticum / Q000276:immunology
D000911:Antibodies, Monoclonal; D012709:Serum Albumin; D013096:Spermine; D010047:Ovalbumin; D013095:Spermidine