INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2024
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IF 7.7
Jie Li,Jikai Gao,Jiaying Ai,Ziyang Yin,Fuping Lu,Hui-Min Qin,Shuhong Mao
17α-Hydroxyprogesterone (17α-OH-PROG) is an important intermediate with a wide range of applications in the pharmaceutical industry. Strategies based on efficient electron transfer and cofactor regeneration were used for the production of 17α-OH-PROG. Here, CYP260A1, Fpr and Adx were expressed using a double plasmid system, resulting in higher biotransformation efficiency. Further optimization of reaction conditions and addition of polymyxin B increased the production of 17α-OH-PROG from 12.52 mg/L to 102.37 mg/L after 12 h of biotransformation. To avoid the addition of external 5-aminolevulinic acid (ALA) as a heme precursor for the P450 enzyme, a modified C5 pathway was introduced into the engineered strain, further reducing the overall process cost. The resulting whole-cell biocatalyst achieved the highest biotransformation yield of 17α-OH-PROG reported to date, offering a promising strategy for commercial application of P450 enzymes in industrial production of hydroxylated intermediates.
研究领域:
生物化学
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制药工程
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合成生物学
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DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
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2024
·
IF 3.8
He Ziyun,Dai Haibing,Shen Jian,Huang Yanjie,Liu Jinsong,Yan Renqing,Zhang Feng,Yan Shengkai
17α-Hydroxyprogesterone (17α-OHP) quantification in dried blood spots (DBS) is essential for newborn screening for congenital adrenal hyperplasia (CAH), which is challenging due to its low physiological concentration. The high false-positive rates of immunoassays necessitate the development of more accurate methods. Liquid chromatography tandem mass spectrometry (LC-MS/MS) offers increased specificity and sensitivity, yet standardized procedures for 17α-OHP measurement are required for clinical application. A candidate reference measurement procedure (cRMP) using isotope dilution LC-MS/MS was developed for 17α-OHP quantification in DBS. By utilizing stable isotope-labeled D8-17α-OHP as an internal standard, the cRMP was optimized, covering sample preparation, calibration, and LC-MS/MS analysis. The method performance was validated across several parameters, including precision, accuracy, specificity, detection limits, and matrix effects. Clinical applicability was further assessed through the establishment of reference intervals for healthy newborns. The developed cRMP exhibited a linear range of 1.00 to 80.00 ng/mL for 17α-OHP, with detection and quantification limits of 0.14 ng/mL and 0.52 ng/mL, respectively. Inter- and intraday precision demonstrated coefficients of variation within 1.27 to 5.69%. The recovery rates and matrix effects were well within acceptable limits, ensuring method reliability. Clinical application showed distinct reference intervals for healthy newborns that were unaffected by sex but influenced by weight and gestational age. This method significantly enhances CAH diagnostic accuracy in newborns, providing a valuable tool for clinical laboratories and improving newborn screening program standardization and traceability.
研究领域:
医学检验
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临床化学
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DOI
PubMed
高影响因子
ANALYTICAL BIOCHEMISTRY
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2020
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IF 2.877
Zhijing Song,Huanyu Gao,Wenchun Xie,Qing Sun,Kai Liang,Yan Li
Measuring the concentrations of steroid hormones in plasma is critical for understanding their role in various vital physiological processes . The detection of underivatized steroid hormones in biofluids through mass spectrometry (MS) is typically hindered by low ionization efficiency. We described a novel matrix-assisted laser desorption/ionization-MS (MALDI-MS) approach based on hydroxylamine derivatization (HA-D) to analyze low-concentration steroid hormones in plasma. The ketonic carbonyl group containing steroid hormones could be derivatized using HA to form oxime derivatives, which considerably enhanced the MS sensitivity for detecting steroid hormones. By using the optimized conditions, estrone (E1), testosterone (T), and progesterone (Prog), could be simultaneously quantified in plasma with a limit of detection (LOD) from 0.019 to 0.031 nM, recoveries from 86% to 108%, and coefficient of variation (CV%) from 4.59% to 11.90%. HA-D/MALDI-MS exhibited higher sensitivity than those using Girard T (GT). To establish potential utility of our method, we characterized fatty liver patient plasmas to demonstrate that the HA-D/MALDI-MS procedure could generate quantitative results comparable to the current clinical liquid chromatography-electrospray ionization tandem MS (LC-ESI MS/MS) method. This approach facilitates the rapid and accurate characterization of plasma hormones , and renders the MALDI-MS approach for steroid hormones more adaptable for clinical research and use.
研究领域:
生物化学
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医学检验
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
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2017
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IF 7.78
Mai Luo,Yifan Hua,Yiran Liang,Jiajun Han,Donghui Liu,Wenting Zhao,Peng Wang
A novel functionalized carbon dot has been synthesized by covalently linking β-cyclodextrin to the surface of N, S codoped carbon dots (β-CD-CDs). The characterization was confirmed by transmission electron microscopy , X-ray photoelectron spectroscopy, infrared spectra, ultraviolet-visible, and fluorescence emission spectra. On the basis of this carbon dot and (ferrocenylmethyl) trimethylammonium iodide (Fc + ), a photo-induced electron transfer (PET) fluorescent probe system was developed to determine the concentration of testosterone in water and identify testosterone in cell by fluorescence imaging as a visible biomarker. Under the optimum condition, the fluorescent intensity of the probe system linearly responded to the concentration of testosterone from 0 μM to 280 μM and the limit of detection was 0.51 μM. This probe system also performed well at determining testosterone in groundwater with average recoveries of testosterone ranging from 96% to 107% at spiking levels of 0.5–100 μM, and the relative standard deviation remained below 13%, which provided a reliable, rapid and easy method to determine testosterone in environmental water. Furthermore, the low cytotoxicity, high anti-interference ability, and excellent biocompatibility of β-CD-CDs made this probe system successfully used in cell fluorescence imaging to monitor levels of testosterone in the cytoplasm of cells with a promising application value in medical research.
研究领域:
分析化学
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环境科学
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生物医学工程
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材料科学
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DOI
PubMed
高影响因子
NEW JOURNAL OF CHEMISTRY
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2018
·Vol. 42
·期 4
·2857-2864
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IF 3.201
Chunxi Zhao,Yang Jiao,Lei Zhang,Yaling Yang
Herein, we report a fast and selective technique for the determination of diethylstilbestrol in milk sample using fluorescent sulfur and boron co-doped carbon dots (S,B co-doped CDs). The S,B co-doped CDs were synthesized by hydrothermal synthesis using poly(sodium-p-styrenesulfonate) and borax as precursors. The as-prepared S,B co-doped CDs possess good stability and fluorescence characteristics and exhibit a fluorescence quantum yield of 25.7%. The fluorescence intensity of the S,B co-doped CDs decreased in the presence of diethylstilbestrol, and a good linear relationship existed between the fluorescence quenching and different concentrations of diethylstilbestrol in the range of 0.1–200 μM with a detection limit of 0.06 μM. Finally, the S,B co-doped CDs were successfully applied in the detection of diethylstilbestrol in milk samples, yielding recoveries between 90.4% and 109.2%.
研究领域:
分析化学
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食品科学
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DOI
高影响因子
危险品化学品经营许可证（带存储）