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Lopatin DE, Winkelhake JL, Voss EW. 
“Immunochemical Characterization of a Lysergyl Derivative Incorporated into Protein”. 
Mol. Pharmacal.. 1974;lO(5):767 - 75.
Abstract
The characterization of the derivative of lysergic acid (LSD; Sandoz) incorporated into rabbit peptide is described, Methods Antibody-producing cells were produced by immunizing rabbits with a fluorescein-conjugated porcin -globulin, then preparing esplenic and/or popliteal lymph node cell cultures. Incubation cultures provided labeled secreted unaltered protein and IBID derivative (Iysergylder) peptides. Anti-lysergyl and anti-lysergylder peptide antisera were produced and antilysergyl reactivity monitored by radioimmunoassay, To reduce peptide components, protease digestion was sometimes used, Alkaline hydrolyses was sometimes used to obtain lysergylder and LSD ligands free of protein, Equilibrium dialysis was performed, Average intrinsic association constants (Ko) were determined from a Scatchard plot. The relative affinities of lysergylder from protease-digested peptides and (3H)LSD for anti-lysergyl and anti-lysergylder antibody were determined by comparative binding radioimmunoassay. Anti-lysergylder-MOPC-315 peptide antibody bound (3H)LSD with à Ko of 8-10 x 10^5 M-l and a heterogeneity index(a) of 0. 74. Anti-lysergylder rabbit peptide antibody bound (3H)LsD with a Ho of 9-10 x l O4M l and a value of 0. 68, Immunization of rabbits with (3H)LSD bound noncovalently to a higher molecular weight serum protein failed to elicit antilysergyl activity, Antilysergyl antibody possessed a Ko of' 3.5 x 10^5 M for (3H)LSD.- Antilysergyl antibody - bound the derivative with a Ko of 7.8 x 1 05M. Comparisons of ligand binding by single-point equilibrium dialyses studies indicate that both antibody populations bound the derivatives in preference to LSD, Alkaline hydrolysis of (3H)LSD and (3H)lysergylder peptides induced no change in antibody binding of lyBergylaer but a 35 0ncrease in binding of (3H)LSD, The hydrolytic product of LSD closely resembled lysergylder.
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