Production and characterization of a rat monoclonal antibody against leu5 enkephalin.
A rat X mouse hybridoma line producing a monoclonal antibody against leu5 enkephalin has been obtained. The monoclonal antibody belongs to the IgG2b class and has an affinity constant of 8.0 X 10(8) M-1 at 4 degrees C. This antibody exhibits approximately 40% cross-reactivity with 1-6 dynorphin but very weak cross-reactivities with met5 enkephalin (1.4%), 1-13 dynorphin (1.3%) and beta-endorphin (0.0045%). The detailed study, by competitive assay, of the interaction between this antibody and various enkephalin derivatives shows that the carboxy-terminal part of the molecule and, particularly, the leu side chain constitutes the immunodominant group. Nevertheless, the tyrosyl residue also contribute considerably to the binding, probably via a peculiar conformation effect, although we can not exclude the tyrosyl residue acting as a secondary contact residue. This monoclonal antibody has been used in a radioimmunoassay to determine leu5 enkephalin like immunoreactive material present in rat brain and hypothalamus.
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D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D000915:Antibody Affinity; D004723:Endorphins / Q000378:metabolism; D004743:Enkephalin, Leucine / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006825:Hybridomas / Q000276:immunology; D011863:Radioimmunoassay; D051381:Rats; D011919:Rats, Inbred Strains; D013329:Structure-Activity Relationship
D000911:Antibodies, Monoclonal; D004723:Endorphins; D004743:Enkephalin, Leucine