Development of an Enzyme-Linked Immunosorbent Assay and Immunoaffinity Column Chromatography for Saikosaponin d Using an Anti-Saikosaponin d Monoclonal Antibody

Sai, Jiayang; Zhao, Yan; Shan, Wenchao; Qu, Baoping; Zhang, Yue; Cheng, Jinjun; Qu, Huihua; Wang, Qingguo

doi:10.1055/s-0035-1568182

Cited by 8 publications

(6 citation statements)

References 34 publications

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“…Several methods for determining saikosaponins in Radix Bupleuri have been described, including thin‐layer chromatography, droplet countercurrent chromatography, micellar electrokinetic capillary chromatography, high‐performance liquid chromatography (HPLC), LC coupled with evaporative light‐scattering detection and charged aerosol detection, LC/MS and immunoassay . Bioanalysis of saikosaponins has also been reported by LC/MS/MS in the biosamples of animals after administration .…”

Section: Introductionmentioning

confidence: 99%

Localization of malonyl and acetyl on substituted saikosaponins according to the full‐scan mass spectra and the fragmentation of sodium‐adduct ions in the positive mode

Liu

Zhang

et al. 2019

Rapid Comm Mass Spectrometry

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Rationale:Discriminating between aglycone-substituted and saccharide-substituted saikosaponins by liquid chromatography/tandem mass spectrometry (LC/MS n ) is a long-standing issue that is still to be resolved. It is necessary to characterize the two types of substituted saikosaponins taking into consideration the potential significant difference in their bioactivity. Methods:Taking the substituents malonyl and acetyl as examples, we developed a MS strategy to discriminate between the aglycone-substituted and saccharidesubstituted saikosaponins through comparing their Y 0 − nH 2 O (n = 1-2) ions from the protonated molecules in the full-scan mass spectra and their B ions in the MS 2 spectra of sodium-adduct molecules in the positive mode. Results:The deprotonated molecules of the aglycone-substituted saikosaponins presented similar fragmentation patterns to those of saccharide-substituted ones in the negative mode, which could not discriminate whether the substitutes were located on the aglycone or the saccharide. In contrast, the Y 0 − nH 2 O (n = 1-2) ions containing or no substituent were observed respectively in the mass fragmentation of the protonated molecules of aglycone-substituted or saccharide-substituted saikosaponins in the positive mode. In addition, the B ions containing or no substituent were observed respectively in the mass fragmentation of the sodiumadduct molecules of the saccharide-substituted or aglycone-substituted saikosaponins in the positive mode. Two aglycone-malonylated saikosaponins were reported for the first time. Conclusions:Whether the substituents were located on the aglycone or the saccharide could be determined according to the Y 0 − nH 2 O (n = 1-2) ions from the protonated molecules in the full-scan mass spectra and the B ions in the MS 2 spectra of sodium-adduct molecules in the positive mode. Our results have updated the mass fragmentation patterns of substituted saikosaponins, which is helpful for the quality control of pharmaceutical preparations containing saikosaponins. More importantly, this MS strategy should be able to be extended to characterize other substituted saponins of bioactive significance in future studies.

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Section: Introductionmentioning

confidence: 99%

Localization of malonyl and acetyl on substituted saikosaponins according to the full‐scan mass spectra and the fragmentation of sodium‐adduct ions in the positive mode

Liu

Zhang

et al. 2019

Rapid Comm Mass Spectrometry

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“…Hybridoma cells that secreted anti-SSd antibodies had been prepared in our team in an earlier study [ 33 ]. These hybridoma cells were injected into BALB/c mice intraperitoneally to produce ascitic fluids.…”

Section: Methodsmentioning

confidence: 99%

“…The titer of anti-SSd MAb was determined by ELISA, and the absorbance was measured at 450 nm by a micro-plate reader. Specificity analyses of MAbs and the purification of IgG were performed according to Sai et al [ 33 ].…”

Section: Methodsmentioning

confidence: 99%

“…By triggering autophagic cell death, SSd can be used to treat drug-resistant and apoptosis-resistant cancers, according to recent studies [ 31 ]. Current standard reference methods for the determination of SSd are mainly based on gas chromatography, liquid chromatography, or mass spectrometry [ 32 , 33 ]. In recent years, novel methods for quality surveys of SSd such as ELISA have been developed.…”

Section: Introductionmentioning

confidence: 99%

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A Highly Sensitive Immunochromatographic Strip Test for Rapid and Quantitative Detection of Saikosaponin d

et al. 2018

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A quantitative lateral-flow immunoassay using gold nanoparticles (AuNPs) conjugated with a monoclonal antibody (MAb) against saikosaponin d (SSd) was developed for the analysis of SSd. The AuNPs were prepared in our laboratory. The AuNPs were polyhedral, with an average diameter of approximately 18 nm. We used the conjugation between AuNPs and MAbs against SSd to prepare immunochromatographic strips (ICSs). For the quantitative experiment, the strips with the test results were scanned using a membrane strip reader, and a detection curve (regression equation, y = −0.113ln(x) + 1.5451, R2 = 0.983), representing the averages of the scanned data, was obtained. This curve was linear from 96 ng/mL to 150 μg/mL, and the IC50 value was 10.39 μg/mL. In this study, we bring the concept of POCT (point-of-care testing) to the measurement of TCM compounds, and this is the first report of quantitative detection of SSd by an ICS.

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“…Saikosaponins are triterpenoids, which are subdivided into saikosaponin a, b1, b2, c and d (10). Saikosaponin a (SSa), Saikosaponin c (SSc) and Saikosaponin d (SSd) exert anti-inflammatory effects (11)(12)(13), and SSa and SSd inhibit cancer cell proliferation and invasion (14)(15)(16). SSa induces apoptosis in McF-7 cells and the process may be regulated by the Bcl-2 family and dependent on p53/p21 signalling pathway (14).…”

Section: Introductionmentioning

confidence: 99%

Antitumor effects of saikosaponin b2 on breast cancer cell proliferation and migration

Gao

et al. 2019

Mol Med Report

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Saikosaponin b2 (SSb2) can be extracted from Bupleurum spp. roots (radix Bupleuri), which belongs to the umbelliferae family. The current study aimed to explore the effects of SSb2 on proliferation of breast cancer cells and to identify the mechanism by which SSb2 affects breast cancer cell migration. mrna expression levels of STaT3 and vasodilator-stimulated phosphoprotein (VaSP) were determined and increased expression was observed in 16 breast cancer tissues compared with the paracancerous tissues. MTT, wound healing, colony formation assays and western blot suggested that SSb2 inhibited McF-7 proliferation and migration. it was further identified by western blot analysis that SSb2 treatment reduced levels of phosphorylated STaT3, VaSP, matrix metallopeptidase (MMP) 2 and MMP9 in McF-7 compared with the untreated cells. in addition, it was demonstrated that inhibition of STaT3 phosphorylation decreased VaSP expression levels and induction of STaT3 phosphorylation increased VaSP levels. Furthermore, it was observed that the treatment of Kunming mice with SSb2 at 30 mg/kg/day for 30 days induced no obvious changes in the liver or kidney tissues, as determined by haematoxylin and eosin staining. in conclusion, these results indicated that SSb2 may be a potential antitumor drug for the treatment of breast cancer, which acts by suppressing proliferation and migration by downregulating the STaT3 signalling pathway and inhibiting the expression of VaSP, MMP2 and MMP9 expression.

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Development of an Enzyme-Linked Immunosorbent Assay and Immunoaffinity Column Chromatography for Saikosaponin d Using an Anti-Saikosaponin d Monoclonal Antibody

Cited by 8 publications

References 34 publications

Localization of malonyl and acetyl on substituted saikosaponins according to the full‐scan mass spectra and the fragmentation of sodium‐adduct ions in the positive mode

Localization of malonyl and acetyl on substituted saikosaponins according to the full‐scan mass spectra and the fragmentation of sodium‐adduct ions in the positive mode

A Highly Sensitive Immunochromatographic Strip Test for Rapid and Quantitative Detection of Saikosaponin d

Antitumor effects of saikosaponin b2 on breast cancer cell proliferation and migration

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