Determination of cross-reactivity of poly- and monoclonal antibodies for synthetic cannabinoids by direct SPR and ELISA

Langer, Nico; Steinicke, Franziska; Lindigkeit, Rainer; Ernst, Ludger; Beuerle, Till

doi:10.1016/j.forsciint.2017.09.011

Cited by 11 publications

(12 citation statements)

References 31 publications

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“…In the more specific field of snake antivenom antibodies, SPR has mainly been used for studying binding kinetics and affinity [ 47 , 48 , 49 , 50 , 51 ], but could also be exploited for the study of antivenom cross-reactivity. Recently, a study by Langer et al [ 52 ], investigating the cross-reactivity of polyclonal and monoclonal antibodies for synthetic cannabinoids, demonstrated the potential of SPR as a valuable tool for this type of investigation due to the reduced amount of manual labor, the real-time analysis, and the high information content obtained about the binding interactions. Also, a newly published study by Choudhury et al [ 53 ] has revealed the development of a portable and inexpensive SPR measurement device with an integrated biosensor for detection of snake venom proteins, and illustrated the use of this device in the study of snake venom protein-antibody interactions.…”

Section: Traditional Studies Of Cross-reactivitymentioning

confidence: 99%

Antibody Cross-Reactivity in Antivenom Research

Ledsgaard

Jenkins

Davidsen

et al. 2018

Toxins

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Antivenom cross-reactivity has been investigated for decades to determine which antivenoms can be used to treat snakebite envenomings from different snake species. Traditionally, the methods used for analyzing cross-reactivity have been immunodiffusion, immunoblotting, enzyme-linked immunosorbent assay (ELISA), enzymatic assays, and in vivo neutralization studies. In recent years, new methods for determination of cross-reactivity have emerged, including surface plasmon resonance, antivenomics, and high-density peptide microarray technology. Antivenomics involves a top-down assessment of the toxin-binding capacities of antivenoms, whereas high-density peptide microarray technology may be harnessed to provide in-depth knowledge on which toxin epitopes are recognized by antivenoms. This review provides an overview of both the classical and new methods used to investigate antivenom cross-reactivity, the advantages and disadvantages of each method, and examples of studies using the methods. A special focus is given to antivenomics and high-density peptide microarray technology as these high-throughput methods have recently been introduced in this field and may enable more detailed assessments of antivenom cross-reactivity.

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Section: Traditional Studies Of Cross-reactivitymentioning

confidence: 99%

Antibody Cross-Reactivity in Antivenom Research

Ledsgaard

Jenkins

Davidsen

et al. 2018

Toxins

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“…The observed cross-reactivities of 6-AmHap-mAb to both 6-AmHap-Cy5 and morHap-Cy5 are not unexpected as these hapten-fluorophore tracers share similar structural faces, which is consistent with the “facial recognition” hypothesis. 38 , 39 …”

Section: Resultsmentioning

confidence: 99%

A Rapid Method for Direct Quantification of Antibody Binding-Site Concentration in Serum

et al. 2022

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The quantitation of the available antibody binding-site concentration of polyclonal antibodies in serum is critical in defining the efficacy of vaccines against substances of abuse. We have conceptualized an equilibrium dialysis (ED)-based approach coupled with fluorimetry (ED-fluorimetry) to measure the antibody binding-site concentration to the ligand in an aqueous environment. The measured binding-site concentrations in monoclonal antibody (mAb) and sera samples from TT-6-AmHap-immunized rats by ED-fluorimetry are in agreement with those determined by a more established equilibrium dialysis coupled with ultraperformance liquid chromatography tandem mass spectrometry (ED-UPLC-MS/MS). Importantly, we have shown that the measured antibody binding-site concentrations to the ligand by ED-fluorimetry were not influenced by the sample serum matrix; thus, this method is valid for determining the binding-site concentration of polyclonal antibodies in sera samples. Further, we have demonstrated that under appropriate analytical conditions, this method resolved the total binding-site concentrations on a nanomolar scale with good accuracy and repeatability within the microliter sample volumes. This simple, rapid, and sample preparation-free approach has the potential to reliably perform quantitative antibody binding-site screening in serum and other more complex biological fluids.

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“…Therefore, the first series of haptens was designed with an alkyl (hapten 1), phenyl (hapten 2), or naphthyl (hapten 3) at the head region. Studies have demonstrated the critical effects of linker position in how the hapten is approached by immune cells. , Thus, in addition to investigating the generation of broadly neutralizing antibodies, haptens 3, 4, 5, 7, and 8 were modified at various regions to understand the effect of linker placement on hapten immunogenicity. In this regard, Class I drugs can be modified at three sites: 4′ naphthyl, 5′ indole, and tail terminal.…”

Section: Results and Discussionmentioning

confidence: 99%

Broadly Neutralizing Synthetic Cannabinoid Vaccines

Lin

Lee

Blake

et al. 2020

JACS Au

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Synthetic cannabinoids (SCs) constitute a significant portion of psychoactive substances forming a major public health risk. Due to the wide variety of SCs, broadly neutralizing antibodies generated by active immunization present an intriguing pathway to combat cannabinoid use disorder. Here, we probed hapten design for antibody affinity and cross reactivity against two classes of SCs. Of the 10 haptens screened, 3 vaccine groups revealed submicromolar IC 50 , each targeting 5–6 compounds in our panel of 22 drugs. Moreover, SCs were successfully sequestered when administered by vaping or intraperitoneal injection, which was confirmed within animal models by observing locomotion, body temperature, and pharmacokinetics. We also discovered synergistic effects to simultaneously blunt two drug classes through an admixture vaccine approach. Collectively, our study provides a comprehensive foundation for the development of vaccines against SCs.

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Determination of cross-reactivity of poly- and monoclonal antibodies for synthetic cannabinoids by direct SPR and ELISA

Cited by 11 publications

References 31 publications

Antibody Cross-Reactivity in Antivenom Research

Antibody Cross-Reactivity in Antivenom Research

A Rapid Method for Direct Quantification of Antibody Binding-Site Concentration in Serum

Broadly Neutralizing Synthetic Cannabinoid Vaccines

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