ArticleHuman vaccines & immunotherapeutics2015
Characterization of a recombinant humanized anti-cocaine monoclonal antibody and its Fab fragment.
Article in Human vaccines & immunotherapeutics, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed, 29 citations in OpenAlex.
- Synthesis and characterization of a functional benzoylecgonine - bovine serum albumin conjugate for quantification of a humanized monoclonal anti-cocaine antibody.Biochemistry and biophysics reports · 2025Article
- Novel partial reduction of the humanized anti-cocaine mAb h2E2 for selective cysteine labeling.Biochemical and biophysical research communications · 2024Article
- Toxicokinetics of a humanized anti-cocaine monoclonal antibody in male and female rats and lack of cross-reactivity.Human vaccines & immunotherapeutics · 2023Review
- Characterization and optimization of fluorescein isothiocyanate labeling of humanized h2E2 anti-cocaine mAb.Biochemistry and biophysics reports · 2023Article
- Isothermal titration calorimetry determination of thermodynamics of binding of cocaine and its metabolites to humanized h2E2 anti-cocaine mAb.Biochemistry and biophysics reports · 2022Article
- Tyrosine nitration of a humanized anti-cocaine mAb differentially affects ligand binding of cocaine and its metabolites.Biochemistry and biophysics reports · 2022Article
- Cocaine binding to the Fab fragment of a humanized anti-cocaine mAb quantitated by dye absorption and fluorescence spectroscopy.Journal of immunological methods · 2021Article
- Ligand binding to a humanized anti-cocaine mAb measured by dye absorption spectroscopy.Biochemical and biophysical research communications · 2021Article
- Multi-domain unfolding of the Fab fragment of a humanized anti-cocaine mAb characterized by non-reducing SDS-PAGE.Biochemical and biophysical research communications · 2020Article
- Ligand binding to a humanized anti-cocaine mAb detected by non-reducing SDS-PAGE.Biochemistry and biophysics reports · 2020Article
- A novel differential scanning fluorimetry analysis of a humanized anti-cocaine mAb and its ligand binding characteristics.Journal of immunological methods · 2020Article
- Structural analysis of free and liganded forms of the Fab fragment of a high-affinity anti-cocaine antibody, h2E2.Acta crystallographica. Section F, Structural biology communications · 2019Article
- Domain unfolding of monoclonal antibody fragments revealed by non-reducing SDS-PAGE.Biochemistry and biophysics reports · 2018Article
- Unfolding of IgG domains detected by non-reducing SDS-PAGE.Biochemical and biophysical research communications · 2018Article
- A mathematical model of a recombinant humanized anti-cocaine monoclonal antibody's effects on cocaine pharmacokinetics in mice.Life sciences · 2017Article
- A novel Python program for implementation of quality control in the ELISA.Journal of immunological methods · 2017Article
- Characterization of a recombinant humanized anti-cocaine monoclonal antibody produced from multiple clones for the selection of a master cell bank candidate.Biochemical and biophysical research communications · 2017Article
- Biologics to treat substance use disorders: Current status and new directions.Human vaccines & immunotherapeutics · 2016Review
- Selective disulfide reduction for labeling and enhancement of Fab antibody fragments.Biochemical and biophysical research communications · 2016Article
- The effects of a repeated dose of a recombinant humanized anti-cocaine monoclonal antibody on cocaine self-administration in rats.Drug and alcohol dependence · 2016Article
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Authors and funding
2 authors at 2 institutions in 1 country.
Funding
Abstract
Variations of post-translational modifications are important for stability and in vivo behavior of therapeutic antibodies. A recombinant humanized anti-cocaine monoclonal antibody (h2E2) was characterized for heterogeneity of N-linked glycosylation and disulfide bonds. In addition, charge heterogeneity, which is partially due to the presence or absence of C-terminal lysine on the heavy chains, was examined. For cocaine overdose therapy, Fab fragments may be therapeutic, and thus, a simplified method of generation, purification, and characterization of the Fab fragment generated by Endoproteinase Lys-C digestion was devised. Both the intact h2E2 antibody and purified Fab fragments were analyzed for their affinities for cocaine and 2 of its metabolites, benzoylecgonine and cocaethylene, by fluorescence quenching of intrinsic antibody tyrosine and tryptophan fluorescence resulting from binding of these drugs. Binding constants obtained from fluorescence quenching measurements are in agreement with recently published radioligand and ELISA binding assays. The dissociation constants determined for the h2E2 monoclonal and its Fab fragment are approximately 1, 5, and 20 nM for cocaethylene, cocaine, and benzoylecgonine, respectively. Tryptophan fluorescence quenching (emission at 330 nm) was measured after either excitation of tyrosine and tryptophan (280 nm) or selective excitation of tryptophan alone (295 nm). More accurate binding constants are obtained using tryptophan selective excitation at 295 nm, likely due to interfering absorption of cocaine and metabolites at 280 nm. These quenching results are consistent with multiple tryptophan and tyrosine residues in or near the predicted binding location of cocaine in a previously published 3-D model of this antibody's variable region.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.