Anti-miroestrol polyclonal antibodies: a comparison of immunogen preparations used to obtain desired antibody properties

Kitisripanya, Tharita; Jutathis, Kamonthip; Inyai, Chadathorn; Komaikul, Jukrapun; Udomsin, Orapin; Yusakul, Gorawit; Tanaka, Hiroyuki; Putalun, Waraporn

doi:10.1007/s11418-015-0949-x

Cited by 9 publications

(12 citation statements)

References 10 publications

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“…Miroestrol‐HSA conjugate prepared according to previous report was used as coating antigen for ELISA described in this study. The reactivity of recombinant anti‐miroestrol scFv antibody against miroestrol‐HSA was evaluated by indirect non‐competitive ELISA.…”

Section: Methodsmentioning

confidence: 99%

“…The developed PAb‐based assays were validated for their high sensitivity and application in determination of miroestrol in P. candollei plant samples and herbal products. We also investigated different methods for preparation of miroestrol immunogen conjugates to obtain PAbs targeting miroestrol with desired properties including improved antibody titer and specificity . These PAbs were obtained from sera of rabbits immunised with miroestrol immunogen conjugates.…”

Section: Introductionmentioning

confidence: 99%

“…We also investigated different methods for preparation of miroestrol immunogen conjugates to obtain PAbs targeting miroestrol with desired properties including improved antibody titer and specificity. 20 These PAbs were obtained from sera of rabbits immunised with miroestrol immunogen conjugates. The process of PAb production is relatively rapid and simple.…”

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confidence: 99%

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Preparation of a highly specific single chain variable fragment antibody targeting miroestrol and its application in quality control of Pueraria candollei by enzyme‐linked immunosorbent assay

Pongkitwitoon

Boonsnongcheep

Kitisripanya

et al. 2019

Phytochemical Analysis

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Introduction Miroestrol is the potent phytoestrogen isolated from White Kwao Krua (Pueraria candollei var. mirifica (Airy Shaw & Suvat.) Niyomdham, a Thai traditional medicinal plant. Nowadays, various health supplementary products featuring White Kwao Krua are available worldwide. A sensitive and rapid analytical method for quantification of miroestrol is necessary for quality control of these products. Objectives To prepare a single‐chain variable fragment (scFv) antibody specific to miroestrol and develop a scFv‐based enzyme‐linked immunosorbent assay (ELISA) for quantitative analysis of miroestrol in plant materials and health supplementary products. Methods A gene encoding anti‐miroestrol scFv antibody was constructed and expressed in Escherichia coli SHuffle T7 strain. Anti‐miroestrol scFv antibody was characterised and applied to ELISA. The developed scFv‐based ELISA method was validated for its sensitivity, specificity, accuracy and precision. Results Anti‐miroestrol scFv antibody was highly specific to miroestrol. The scFv‐based ELISA was applied to determine miroestrol in the range 0.06–7.81 μg/mL, with the limit of quantification of 0.06 μg/mL miroestrol. The accuracy of the assay was validated by its 95.08–103.99% recovery from the spiked miroestrol recovery experiment and in good correlation with the results from the monoclonal antibody‐based ELISA. The relative standard deviation of the intra‐ and inter‐assay were less than 6.0%. Conclusion The developed scFv‐based ELISA was sensitive, specific, accurate, and precise for determination of miroestrol and useful for quality control of P. candollei plant raw materials and supplementary products.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Preparation of a highly specific single chain variable fragment antibody targeting miroestrol and its application in quality control of Pueraria candollei by enzyme‐linked immunosorbent assay

Pongkitwitoon

Boonsnongcheep

Kitisripanya

et al. 2019

Phytochemical Analysis

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“…Miroestrol analysis: Indirect competitive enzyme-linked immunosorbent assay method using anti-miroestrol polyclonal antibody was used to determine miroestrol quantitation [28]. The 96-well ELISA plates were coated with 100 µL of 5 µg/mL miroestrol-cOVA conjugate solution prepared in 50 mM carbonate buffer (pH 9.6) and incubated for 1 h at 37 °C then it was washed three times with TPBS.…”

Section: Deoxymiroestrol Analysismentioning

confidence: 99%

Isoflavonoids and Phytoestrogens from Pueraria candollei var. mirifica Related with Appropriate Ratios of Ethanol Extraction

Peerakam¹,

Sirisa-ard²,

Huy³

et al. 2018

Asian J. Chem.

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“…The developed immunoassay showed high specificity and sensitivity against miroestrol, and sample preparation was relatively simple. Furthermore, we have investigated immunogen preparation to obtain an antibody against miroestrol with the desired characteristics, including improved specificity and antibody titer [21]. In the current study, a monoclonal antibody (mAb) against miroestrol was now generated to provide a sustainable source of antibody for immunoassay development.…”

Section: Development Of An Enzyme-linked Immunosorbent Assay For Detementioning

confidence: 99%

Development of an Enzyme-Linked Immunosorbent Assay for Determination Of Miroestrol Using an Anti-miroestrol Monoclonal Antibody

et al. 2017

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Miroestrol is a chromene with potent estrogenic activity present in , commonly known as White Kwao Krua. Although this compound is only present in low amounts in the plant, it plays an important role in the estrogenic action of products. As a tool for further studies about the efficacy and safety of as a phytoestrogenic supplement, we generated a novel monoclonal antibody against miroestrol. This anti-miroestrol monoclonal antibody was used to develop an immunoassay for the determination of miroestrol content, which can be used for quality control purposes of. The developed ELISA against miroestrol has a calibration range of 10-780 ng/mL miroestrol, a limit of detection of 3.5 ng/mL, and a limit of quantitation of 12.2 ng/mL. According to the validation analysis, the established ELISA is precise, accurate, specific, and sensitive for miroestrol detection in plants. Furthermore, the anti-miroestrol monoclonal antibody was used to prepare an immunoaffinity column for the isolation of miroestrol from the tuberous root of . The column provides a simple procedure for miroestrol isolation, with a capacity of 3.91 µg of miroestrol per 1 mL of immunogel.

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Anti-miroestrol polyclonal antibodies: a comparison of immunogen preparations used to obtain desired antibody properties

Cited by 9 publications

References 10 publications

Preparation of a highly specific single chain variable fragment antibody targeting miroestrol and its application in quality control of Pueraria candollei by enzyme‐linked immunosorbent assay

Preparation of a highly specific single chain variable fragment antibody targeting miroestrol and its application in quality control of Pueraria candollei by enzyme‐linked immunosorbent assay

Isoflavonoids and Phytoestrogens from Pueraria candollei var. mirifica Related with Appropriate Ratios of Ethanol Extraction

Development of an Enzyme-Linked Immunosorbent Assay for Determination Of Miroestrol Using an Anti-miroestrol Monoclonal Antibody

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