Abstract
When generating monoclonal antibodies (mAb) against small molecules, the chemical composition and molecular orientation of the drug-like hapten on the antigen is a crucial determinant. This is especially important when attempting to discover therapeutic mAb against the drugs of abuse (+)-methamphetamine [(+)-METH], (+)-amphetamine [(+)-AMP], and the related compound (+)-3,4-methylenedioxymethamphetamine [(+)-MDMA, the plus isomer in the racemic mixture known as MDMA or ecstasy]. The goal of these studies was to design and synthesize (+)-METH-like haptens with structural attributes that could make them effective for generating monoclonal antibodies for treating medical problems associated with these stimulant drugs of abuse. Five prototype (+)-METH-like haptens, which mimic structural aspects of these drugs, were synthesized and used to generate mAb. After screening for anti-(+)-METH IgG antibodies in more than 25,000 potential mouse hybridoma cell lines, one prototype mAb from each of the five haptens was selected and studied in detail for molecular properties and preclinical efficacy. The amino acid sequences of the IgG-variable regions, structural models, affinity, and ligand specificity of each mAb were then used to help elucidate important therapeutic characteristics. Four of these antibodies exhibited high affinity and specificity to (+)-METH and (+)-MDMA; whereas one antibody (designated mAb4G9) exhibited high affinity and specificity to (+)-METH, (+)-MDMA, and (+)-AMP, without significant cross-reactivity against other METH-like ligands, over-the-counter medications, or endogenous neurotransmitters. Considered together, discovery of mAb4G9 and the other antibodies in this report represent an important step in understanding the process for custom design of drug class-specific therapeutic antibodies for the treatment of drug addiction.
Footnotes
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This work was supported by National Institutes on Drug Abuse Grants R01 DA11560, P01 DA14361, F32 DA018039, and R37 DA05477 and National Institutes of Health Centers of Biomedical Research Excellence Grant P20 RR15569. S.M.O. and R.H. have financial interests in and serve as Chief Scientific Officer and Vice President of Pharmaceuticals, respectively, of InterveXion Therapeutics LLC (Little Rock, AR), a pharmaceutical biotechnology company whose main interest is in developing new monoclonal antibodies for treatment of human diseases, including drug abuse.
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Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.
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doi:10.1124/jpet.106.117150.
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ABBREVIATIONS: (+)-METH, (+)-methamphetamine; mAb, monoclonal antibody(ies); (+)-AMP, (+)-amphetamine; (+)-MDMA, (+)-3,4-methylenedioxymethamphetamine; (–)-METH, (–)-methamphetamine; (–)-AMP, (–)-amphetamine; (–)-MDMA, (–)-3,4-methylenedioxymethamphetamine; BSA, bovine serum albumin; RIA, radioimmunoassay; WAM, web antibody modeling; CDR, complementary-determining region(s); RMSD, root mean square distance.
- Received November 16, 2006.
- Accepted February 5, 2007.
- The American Society for Pharmacology and Experimental Therapeutics
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