Ultrasensitive immunoassay for monocrotaline using monoclonal antibody produced by N, N’ -carbonyldiimidazole mediated hapten-carrier protein conjugates

Sakamoto, Seiichi; Nagamitsu, Rika; Yusakul, Gorawit; Miyamoto, Tomofumi; Tanaka, Hiroyuki; Morimoto, Satoshi

doi:10.1016/j.talanta.2017.03.028

Cited by 24 publications

(23 citation statements)

References 22 publications

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“…Previously, the MAb 4B8‐based icELISA was evaluated for its binding specificity against MCT and its related DHPAs . The results suggested that the MAb 4B8‐based icELISA showed binding specificity towards MCT, where CRs were evaluated by the method described previously .…”

Section: Resultsmentioning

confidence: 93%

“…Using the icELISA procedure, the half‐maximum inhibitory concentration (IC 50 ) of each compound was calculated. The CR was then obtained using the reported equation . The IC 50 values of MCT and RN were 0.242 and 230 ng/mL, respectively.…”

Section: Resultsmentioning

confidence: 99%

“…The monocrotaline (MCT) concentrations of Crotalaria spp. extracts were determined by MAb 4B8‐based icELISA …”

Section: Resultsmentioning

confidence: 99%

“…According to the specific binding profile for MAb 4B8, which was determined via icELISA, the MAb 4B8 specificity in PI‐ICA was recharacterised using different DHPAs (i.e. RN, retrorsine, senecionine, and heliotrine) and DHPA‐free plant samples ( Glycyrrhiza glabra , Glycine max , Pueraria mirifica , P. lobata , and Coleus forskohlii ).…”

Section: Methodsmentioning

confidence: 99%

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Preincubation format for a sensitive immunochromatographic assay for monocrotaline, a toxic pyrrolizidine alkaloid

Yusakul

Sakamoto

Chanpokapaiboon

et al. 2019

Phytochemical Analysis

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Introduction Monocrotaline (MCT), which is classified as a 1,2‐dehydropyrrolizidine alkaloid (DHPA), is a toxic compound that is mainly produced by Crotalaria spp. MCT contamination in cereals and herbs leads to hepatitis, gastroenteritis, pulmonary vasculitis and hypertension, and different types of cancer. The current analytical methods for MCT are complicated and expensive using liquid chromatography equipped with mass spectrometry detection. Objective The aim of this study was to develop a simple and sensitive preincubation format for an immunochromatographic assay (PI‐ICA) for MCT detection. Methodology We conducted the PI‐ICA via incubation of an MCT‐containing sample with an anti‐MCT monoclonal antibody conjugated with colloidal gold before strip dipping. We compared the PI‐ICA detection sensitivity with that of the conventional ICA (Conv‐ICA) format. Results The PI‐ICA was sensitive with a limit of detection (LOD) of 0.61 ng/mL, which is a 16‐fold improvement over the Conv‐ICA format. These results indicated that the PI‐ICA method exhibits high binding specificity for MCT and low cross‐reactivity towards retronecine, retrorsine, senecionine and heliotrine. Sample solutions from plants containing MCT and related DHPAs produced positive results via PI‐ICA analysis. Conclusions The proposed PI‐ICA system provides a highly sensitive method compared to Conv‐ICA. In addition, the developed PI‐ICA method is simple and highly effective for detecting MCT contamination.

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Section: Resultsmentioning

confidence: 93%

Section: Resultsmentioning

confidence: 99%

“…The monocrotaline (MCT) concentrations of Crotalaria spp. extracts were determined by MAb 4B8‐based icELISA …”

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Preincubation format for a sensitive immunochromatographic assay for monocrotaline, a toxic pyrrolizidine alkaloid

Yusakul

Sakamoto

Chanpokapaiboon

et al. 2019

Phytochemical Analysis

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“…Considering our recent reports about specic MAbs prepared using BSA conjugates containing only two target molecules, [20][21][22] very low hapten numbers may be sufficient for producing highly specic antibodies although some study concluded between 8 and 25 hapten number were optimal.…”

Section: Discussionmentioning

confidence: 99%

Sensitive quantitative analysis of the bitter glycoside amarogentin by specific monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay

et al. 2018

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Amarogentin (AG) is a naturally occurring secoiridoid glycoside produced mainly in the plant genera Swertia and Gentiana. Extracts of these plants have a long history of use in Japan as bitter stomachics because of their strong bitterness. Because the AG content directly reflects the potential activity of the extract, the quality control of these plant extracts through the quantitative analysis of AG is important. In the present study, we aimed to develop a quantitative analysis of AG using a monoclonal antibody (MAb) against AG (MAb 1E9) in the plant family Gentianaceae. Surprisingly, production of MAb 1E9 was successfully achieved by immunizing AG-bovine serum albumin (BSA) conjugates into mice although the number of AG bound to BSA was only one. The characterization of MAb 1E9 revealed that it shows high specificity to AG, enabling the development of an icELISA for the specific determination of AG. In addition, the detectable concentration of AG in the developed icELISA ranged from 1.95 to 62.5 ng mL À1 with a limit of detection of 1.28 ng mL À1, which is approximately 30-625 times higher in sensitivity compared with the conventional HPLC method. Validation analysis revealed that the icELISA using MAb 1E9 is precise (intra-assay variation <3.9%, inter-assay variation <8.8%) and accurate (recovery rates of spiked samples were between 91.0% and 106.4%). This method can be used for the quality control of plant extracts using AG as an index.

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A monoclonal antibody‐based indirect competitive enzyme‐linked immunosorbent assay to quantify swertiamarin and related compounds in Swertia japonica Makino

Nuntawong

Horikawa

Ochi

et al. 2020

Phytochemical Analysis

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Introduction: Swertia japonica Makino (S. japonica) has a long history of use as a folk medicine, and it is one of the three essential Japanese folk medicines. S.japonica has been reported to have various biological activities. The biologically active secoiridoid glycoside swertiamarin (SM) has been isolated from S. japonica. The efficacy of this plant is attributed to SM and related secoiridoid glycosides. To control the quality of S. japonica for medicinal use, a method for the determination of SM and other secoiridoid glycosides in the plant is needed.Objective: To produce an anti-SM monoclonal antibody (MAb) and develop an indirect competitive enzyme-linked immunosorbent assay (icELISA) for S. japonica standardisation and quality control.Methodology: SM was conjugated to cationised bovine serum albumin (cBSA), and the SM-cBSA conjugate was used to immunise BALB/c mice. Splenocytes from the immunised mice were then fused with SP2/0 myeloma cells to produce hybridoma cells that expressed anti-SM MAb.Results: The developed icELISA was sufficiently sensitive and had a quantitative range of 0.78 to 12.5 μg/mL. Coefficients of variation below 10% indicated good repeatability. Recoveries in a spike and recovery assay ranged from 91.84% to 115.50%, which confirmed that the icELISA was accurate. The SM content measured using the icELISA was in agreement with the results of a high-performance liquid chromatography-ultraviolet (HPLC-UV) assay. Conclusion:The icELISA is suitable for the high-throughput analysis of SM and other secoiridoid glycosides in S. japonica. The method is fast, economical, and reliable for S. japonica quality control.

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Ultrasensitive immunoassay for monocrotaline using monoclonal antibody produced by N, N’ -carbonyldiimidazole mediated hapten-carrier protein conjugates

Cited by 24 publications

References 22 publications

Preincubation format for a sensitive immunochromatographic assay for monocrotaline, a toxic pyrrolizidine alkaloid

Preincubation format for a sensitive immunochromatographic assay for monocrotaline, a toxic pyrrolizidine alkaloid

Sensitive quantitative analysis of the bitter glycoside amarogentin by specific monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay

A monoclonal antibody‐based indirect competitive enzyme‐linked immunosorbent assay to quantify swertiamarin and related compounds in Swertia japonica Makino

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