Monoclonal antibodies to a nitroxide lipid hapten |
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Authors: | Krishna Balakrishnan Frank J. Hsu Dean G. Hafeman Harden M. McConnell |
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Affiliation: | Stauffer Laboratory for Physical Chemistry, Stanford University, Stanford, CA 94305 U.S.A. |
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Abstract: | The isolation and characterization of a hybridoma cell line producing a monoclonal IgG1 antibody against a spin-label nitroxide group is described. The antibody recognizes a synthetic hapten containing linked dinitrophenyl and 2,2,6,6-tetramethylpiperidinyl 1-oxy groups, having an affinity of 3.6±1.0·106 M?1 for the soluble hapten at 25°C. The antibody binds to phospholipid vesicles containing 2 mol% of spin label-derivitized lipid (lipid hapten) with an affinity of 1.5±0.2·108 M?1. This monoclonal IgG1 mediates the binding of hapten-bearing lipid vesicles to mouse macrophage RAW264 cells bearing Fc receptors. The cellular responses to this binding are similar to those observed previously using polyclonal rabbit anti-hapten IgG. As with the heterogeneous antibodies, the monoclonal IgG1 is more efficient in mediating cellular uptake when the vesicles are in the ‘fluid’ physical state (dimyristoylphosphatidylcholine at 37°C) compared to ‘solid’ (dipalmitoylphosphatidylcholine at 37°C). Despite the enhanced binding of ‘fluid’ phospholipid vesicles to cells, only the ‘solid’ vesicles triggered a significant respiratory burst in RAW264 macrophages. |
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Keywords: | Spin label Nitroxide lipid hapten (Hybridoma cell) TEMPO 2,2,6,6-tetramethyl piperidinyl 1-oxy DPPE dipalmitoylphosphatidylethanolamine DMPC dimyristoylphosphatidylcholine DPPC dipalmitoylphosphatidylcholine |
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