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Development of a plant-produced recombinant monoclonal antibody against Δ-9-tetrahydrocannabinol (Δ9-THC) for immunoassay application.

ABSTRACT: Δ-9-tetrahydrocannabinol (Δ9-THC) is mainly a psychoactive compound in the cannabis plant. The immunoassay, an alternative method to HPLC and GC, can be used to analyze and measure Δ9-THC. This method provides high sensitivity and specificity by using antibodies specific to the desired substances. Currently, plants provide several benefits over traditional expression platforms to produce recombinant antibodies, such as lower production costs and scalability. Therefore, this study aims to produce a recombinant anti-Δ9-THC monoclonal antibody (mAb) with transient expression using N. benthamiana. The highest expression level of the plant-produced mAb was estimated to be 0.33 ug/g leaf fresh weight. Our results demonstrate that the antibody provided in vitro affinity binding related to Δ9-THC and the metabolites of Δ9-THC, such as cannabinol (CBN). Moreover, the antibody also showed binding efficiency with Δ9-THC in cannabis extract. Moreover, plant-produced mAbs provide efficiency against Δ9-THC and can be applied for further immunoassay applications.

SUBMITTER: Boonyayothin W

PROVIDER: S-EPMC9171438 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature

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Development of a plant-produced recombinant monoclonal antibody against Δ-9-tetrahydrocannabinol (Δ9-THC) for immunoassay application.

Boonyayothin Wanuttha W Kobtrakul Khwanlada K Khositanon Petlada P Vimolmangkang Sornkanok S Phoolcharoen Waranyoo W

Biotechnology reports (Amsterdam, Netherlands) 20220326

Δ-9-tetrahydrocannabinol (Δ9-THC) is mainly a psychoactive compound in the cannabis plant. The immunoassay, an alternative method to HPLC and GC, can be used to analyze and measure Δ9-THC. This method provides high sensitivity and specificity by using antibodies specific to the desired substances. Currently, plants provide several benefits over traditional expression platforms to produce recombinant antibodies, such as lower production costs and scalability. Therefore, this study aims to produce ...[more]

PMID: 35686006

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Project description:Delta-9-tetrahydrocannabinol (Δ9-THC) is a naturally occurring psychoactive compound derived from the hemp plant Cannabis sativa. In 2015, EFSA established an acute reference dose (ARfD) of 1 μg/kg body weight (bw) for Δ9-THC and assessed acute dietary exposure from milk and dairy products. This resulted at the most 3% and 13% of the ARfD for adults and toddlers, respectively. Following the European Commission Recommendation 2016/2115 on the monitoring of the presence of Δ9-THC in food and the issuing of a new mandate, EFSA assessed the acute human exposure to Δ9-THC. 'Standard' food categories were used as proxies for consumption of hemp and hemp-based products. Twelve independent scenarios based on single food categories were considered and acute exposure was assessed for consumption days only for all age groups excluding infants. Occurrence data for Total-Δ9-THC (588 samples in total) were used for this assessment up to the highest reliable percentile for each food category. The EFSA ARfD of 1 μg/kg bw was exceeded in the adult high consumers of most considered hemp and hemp-containing products, under the lower-bound (LB) and upper-bound (UB) scenario. At the UB, acute exposure in adult high consumers was estimated based on the highest reliable percentile of occurrence, for Hemp seeds (P95, up to 9 μg/kg bw), Hemp oil (P95, up to 21 μg/kg bw), Tea (Infusion) (P95, up to 208 μg/kg bw), Breakfast cereals (P50, up to 1.3 μg/kg bw), Pasta (Raw) (P75, up to 6.4 μg/kg bw), Bread and rolls (P75, up to 1.3 μg/kg bw), Bread and rolls from hemp flour (P90, up to 4.1 μg/kg bw), Cereal bars (P50, up to 0.3 μg/kg bw), Fine bakery wares (P75, up to 5.1 μg/kg bw), Chocolate (Cocoa) products (P75, up to 1.1 μg/kg bw), Energy drinks (P75, up to 0.2 μg/kg bw), Dietary supplements (P75, up to 9.9 μg/kg bw), Beer and beer-like beverages (P90, up to 41 μg/kg bw). The use of proxies for the consumption of hemp and hemp-containing products, the limited number of occurrence data and the analytical limitations in the quantification of Δ9-THC represent the most important sources of uncertainty. Overall, exposure estimates presented in this report are expected to represent an overestimation of acute exposure to Δ9-THC in the EU.

Project description:BackgroundCannabis is the most common illicit drug used in the USA and its use has been rising over the past decade, while the historical gap in rates of use between men and women has been decreasing. Sex differences in the effects of cannabinoids have been reported in animal models, but human studies are sparse and inconsistent. We investigated the sex differences in the acute subjective, psychotomimetic, cognitive, and physiological effects of intravenous (IV) delta-9 tetrahydrocannabinol (THC), the main psychoactive constituent of cannabis.MethodsHealthy male and female individuals, with limited exposure to cannabis, participated in a double blind, placebo-controlled study of intravenous (IV) placebo or THC at two doses (0.015 mg/kg and 0.03 mg/kg). Visual analog scale (VAS) was used to measure subjective effects, Psychotomimetic States Inventory (PSI) and the Clinician-Administered Dissociative Symptoms Scale (CADSS) were used to assess the psychotomimetic effects and perceptual alterations, respectively, and Rey Auditory Verbal Learning Task (RAVLT) was used to evaluate cognitive effects. Outcome variables were represented as the peak change from baseline for each variable, except RAVLT which was used only once per the test day after the subjective effects.ResultsA total of 42 individuals participated in this study. There were no significant differences between male and female participants in background characteristics. There was a significant main effect of sex on the VAS scores for THC-induced "High" (F1,38 = 4.27, p < 0.05) and a significant dose × sex interaction (F2,77 = 3.38, p < 0.05) with female participants having greater "High" scores than male participants at the lower THC dose (0.015 mg/kg). No other sex differences were observed in acute subjective, psychotomimetic, cognitive, or physiological effects of THC.ConclusionThere were significant sex differences in subjective effects of feeling "High" at a lower dose of THC. However, there were no other sex-related differences in the subjective, physiological, or cognitive effects of THC.

Dataset Information

Development of a plant-produced recombinant monoclonal antibody against Δ-9-tetrahydrocannabinol (Δ9-THC) for immunoassay application.

Publications

Development of a plant-produced recombinant monoclonal antibody against Δ-9-tetrahydrocannabinol (Δ9-THC) for immunoassay application.

Similar Datasets

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