Abstract
Molecular complexes of four monoclonal anti-morphine antibodies (mAb) with the opiate ligands morphine, oxymorphone, and naloxone were studied using UV-VIS absorption spectroscopy. Although strong overlaps in the absorption spectra of the antibodies, ligands, and complexes were observed, a curve-fitting method was developed to correlate the absorbance with the concentration of the ligand-antibody complex. Using this technique, we determined the intrinsic association constants for the mAb with morphine to be in the nanomolar range, while association constants for oxymorphone and naloxone were in the micromolar range. These values were found to be in agreement with previous radioimmunoassay determinations. We also observed different changes in the absorbancy of the mAb upon complexation with different ligands and such changes were found to be different for all four mAb examined. Upon complexation with the ligand morphine, two of the mAb (clone numbers MOR368-21 and MOR10.5) displayed distinct charge-transfer spectral bands in the 320-nm region. These observations suggest that mAb binding site tryptophans may participate in the formation of the antibody-ligand complex and such complexation involves a charge-transfer interaction.
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Abbreviations
- CAMM:
-
computer-aided molecular modeling
- CDR:
-
complementarity-determining region
- corr:
-
correlation coefficient
- K A :
-
assocation constant
- mAb:
-
monoclonal antibody
- PBS:
-
phosphate buffered saline,pH 7.0.
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Droupadi, P.R., Meyers, E.A. & Linthicum, D.S. Spectroscopic evidence for charge-transfer complexation in monoclonal antibodies that bind opiates. J Protein Chem 13, 297–306 (1994). https://doi.org/10.1007/BF01901562
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DOI: https://doi.org/10.1007/BF01901562