Nanoscale
·
2024
·
IF 5.8
Xinlv Peng,Caixia Pei,Eika W. Qian,Yuguang Du,Jian-Jun Li
6′-Sialyllactose (6′-SL), the most abundant sialylated human milk oligosaccharide, has attracted attention for its potential application in supplementary infant formulas. Herein, we report a facile strategy to construct a cascade bioreactor for the enzymatic synthesis of 6′-SL by co-immobilizing an enzymatic module consisting of CMP–sialic acid synthase and α-2,6-sialyltransferase into hierarchically porous MIL-53 (HP-MIL-53). The as-prepared HP-MIL-53 showed high enzyme immobilization capacity, reaching 226 mg g−1. Furthermore, the co-immobilized enzymes exhibited higher initial catalytic efficiency, and thermal, pH and storage stability than the free ones. Finally, the 6′-SL yield remained >80% after 13 cycles of use. We expect that HP-MIL-53 would have potential industrial applications in the enzymatic modular synthesis of 6′-SL and other glycans.
研究领域:
生物化学
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食品科学
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材料科学
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DOI
PubMed
高影响因子
Molecular Omics
·
2021
·Vol. 17
·期 4
·565-571
·
IF 3.743
Lujie Yang,Quanqing Zhang,Ling Lin,Ying Xu,Yuanyu Huang,Zuojian Hu,Ke Wang,Cuiping Zhang,Pengyuan Yang,Hongxiu Yu
Glycosylation is a widely occurring posttranslational modification. Here, we applied a quick, convenient and high-throughput strategy (lectin array) to investigate the variation in glycans on different macrophage subtypes derived from THP-1 and RAW264.7 cells. For THP-1 cells, there were more significant differences in the glycan on M2 macrophages compared to the other two subtypes. In contrast, M1 macrophages exhibited more significant glycan remodeling than the other subtypes for the RAW264.7 cell line. The response of the lectins which recogonize the N-glycan and α2,6 sialic acid was higher during polarization into anti-inflammatory phase (THP-1 derived M2 subtypes), and lower in pro-inflammatory phase (RAW264.7 M1 subtypes). The regulation of several α2,6 sialyltransferase genes was coincident with the regulation of the α2,6 sialic acid on the two cell lines. The lectin response and glycosyltranferase gene expression confirmed that α2,6 sialic acid showed higher expression in the anti-inflammatory phase. This indicated that α2,6 sialic acid was a potential indicator for the anti-inflammatory response.
研究领域:
免疫学
·
分子生物学
·
细胞生物学
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2015
·
IF 6.409
Yuliang Li,Junlin He,Yazhen Niu,Chao Yu
In this paper, a simple, ultrasensitive and label-free electrochemical α2,6-sialylated glycans biosensor based on reduced graphene oxide-tetraethylene pentamine-1-butyl-3-methylimidazolium hexafluorophosphate (BMIMPF 6 ) hybrids was developed. Due to the abundance of amino groups from reduced graphene oxide-tetraethylene pentamine (rGO-TEPA) and the electrostatic interaction of BMIMPF 6 , bimetallic gold platinum alloy nanoparticles (AuPtNPs) were densely adsorbed onto the surface of the nanocomposite , providing a large surface area available for the immobilization of Sambucus nigra agglutinin (SNA). AuPtNPs have excellent conductivity and catalytic activity , which can promote electron transfer between the electrolyte solution and the surface of electrode and can enhance the sensitivity of biosensor. SNA, which specifically binds α2,6-sialylated glycans, was covalently immobilized on AuPtNPs for specific detection of α2,6-sialylated glycans in human serum. Under optimal experimental conditions, amperometric response changes were used to detect α2,6-sialylated glycans with a broad linear range of 10 fg mL −1 –1 μg mL − 1 and a low detection limit of 3 fg mL − 1 ( S / N =3). When applied to spiked serum samples, the recovery of the developed biosensor ranged from 100.8% to 101.4%, suggesting that the electrochemical biosensor would be suitable for the practical detection of α2,6-sialylated glycans.
研究领域:
生物传感技术
·
医学诊断
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DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2020
·
IF 5.977
Qihao Zhu,Weiran Mao,Chengli Zhang,Yuan Zhou,Zhiyong Tang,Chao Yu
Here, a novel electrochemiluminescence biosensor based on potential-resolved strategy was firstly prepared for the detection of dual targets α2,3-sialylated glycans and α2,6-sialylated glycans. This is the first time that Au@BSA microsphere was used to connect with luminol to enhance its ECL intensity, and it can generate ECL signals at positive potential. Zeolitic Imidazolate Framework-8 (ZIF-8) and Meso-tetra (4-carboxyphenyl) porphyrin (TCPP) were linked using a one-pot method to synthesize a novel luminescent ZIF (L-ZIF) named TZZ, which can emit ECL signals at negative potential. Moreover, magnetite microspheres were used to construct a sandwich-type biosensor to obtain higher sensitivity and reduce background signals. In addition, the biosensor manufactured directly in solution have a wider linear range than constructed on electrode because it has more available space than the electrode surface. Due to the above advantages, the prepared ECL biosensor exhibited high sensitivity, stability and broader linear range, even for practical analysis. Therefore, the prepared ECL biosensor will become a promising method for determination of α2,3-sialylated glycans and α2,6-sialylated glycans in clinical applications in the future. What is more, it provides a potential method for detection of other multi-targets.
研究领域:
分析化学
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生物医学工程
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传感技术
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2014
·
IF 6.451
Liuliu Gao,Junlin He,Wailan Xu,Jing Zhang,Junmin Hui,Yanlei Guo,Wenjuan Li,Chao Yu
α2,6-Sialylated glycans are crucial molecular targets for cancer diagnosis and clinical research. In this work, a novel ultrasensitive electrochemical biosensor was fabricated based on a graphite oxide (GO), Prussian blue (PB), and PTC-NH 2 (an ammonolysis product of 3,4,9,10-perylenetetracarboxylic dianhydride) nanocomposite for the selective detection of α2,6-sialylated glycans. To increase the sensitivity of the electrochemical biosensor, gold nanoparticles (GNPs) were immobilized on a GO–PB–PTC-NH 2 modified glassy carbon electrode (GCE). Sambucus nigra agglutinins (SNAs), which specifically bind with α2,6-sialylated glycans, were covalently immobilized on GNPs for the sensitive detection of α2,6-sialylated glycans in serum. This proposed method can be applied to human serum, and it worked well over a broad linear range (0.1 pg mL −1 –500 ng mL −1 ) with detection limits of 0.03 pg mL −1 . Moreover, recovery of the spiked samples ranged from 100.2% to 105.0%, suggesting that this excellent electrochemical biosensor can be used for the practical detection of α2,6-sialylated glycans.
研究领域:
生物医学工程
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分析化学
·
癌症诊断
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DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2020
·Vol. 187
·期 12
·1-9
·
IF 6.232
Zhao Yunfei,Chen Jun,Zhong Hangtian,Zhang Chengli,Zhou Yuan,Mao Weiran,Yu Chao
The development of a novel signal amplification system is described for sensitive determination of α2,6-sialylated glycans (α2,6-sial-Gs), an important prognostic tumor biomarker. First, Fe-based metal-organic frameworks (Fe-MOFs) with silver nanoparticles (AgNPs) decorated onto the outer surface were designed and synthesized with controlled octahedron structures. The new Ag/Fe-MOFs nanocomposite possessed strong conductivity and a large surface area to carry more nanoprobes. To connect the Ag/Fe-MOFs nanocomposite with more groups, the nanocomposite was functionalized by -COOH with SH-PEG-COOH to bind with an α2,6-sial-Gs catcher, M-APBA, via -CONH- bonds. More importantly, the Ag/Fe-MOFs also exhibited an excellent endogenous redox mediator property to produce electrons, which is the fundamental mechanism underlying amplification of an electronic signal. A gold electrode was used to accelerate electron transfer and immobilize the α2,6-sial-Gs lectin (SNA). After the sandwich-type catcher recognition (SNA/α2,6-sial-Gs/M-APBA), the current peak response was provoked in the process of oxidizing AgNPs to Ag + in the forward anodic potential sweep, while Cl − in a PBS solution was transferred into Ag + to maintain charge neutrality. Optimized particles were employed for direct fabrication of the sandwich-type affinity biosensor, which was found to show a linear detection range from 1 fg mL −1 to 1 ng mL −1 with a detection limit of 0.09 fg mL −1 . Furthermore, the biosensor exhibited excellent specificity and stability, indicating that such a novel nanobiotechnology platform can be used to initiate potential utility for monitoring biomarkers in serum. Graphical abstract
(A)Schematic presentation of synthesis and surface modification of Ag/Fe-MOFs. The new Ag/Fe-MOFs nanocomposite possessed commendable conductivity and large surface area to carry more nanoprobe; after functionalizing the Ag/MOFs with SH-PEG-COOH, the functionalized endogenous redox mediator (c-Ag/MOFs) realized the possibility that can connect with the biological catcher. (B) Schematic diagram of electrode construction for detecting α2,6-sialylated glycans (α2,6-sial-Gs). By using the c-Ag/Fe-MOFs functional endogenous redox mediator, we successfully implemented the electrochemical detection of α2,6-sial-Gs.
研究领域:
分析化学
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生物材料
·
生物技术
·
电化学
·
癌症诊断
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产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2019
·
IF 9.518
Jia Li,Junlin He,Chengli Zhang,Jun Chen,Weiran Mao,Chao Yu
In this study, a dual-type responsive electrochemical biosensor was developed for the quantitative detection of α2,6-sialylated glycans (α2,6-sial-Gs), a potential biomarker of tumors. The gold nanorods (AuNRs), which exhibited great specific surface area, as well as good biocompatibility , was synthesized by the way of seed growth method. Furthermore, a biotin-streptavidin (biotin-SA) system was introduced to improve the immunoreaction efficiency. Accordingly, a label-free biosensor was fabricated based on AuNRs-SA for the quick detection of α2,6-sial-Gs by recording the signal of differential pulse voltammetry (DPV). Furthermore, to expand the ultrasensitive detection of α2,6-sial-Gs, a carboxylated single-walled carbon nanohorns/sulfur-doped platinum nanocluster (c-SWCNHs/S-PtNC) was synthesized for the first time as a novel signal label, which showed an excellent catalytic performance. The usage of c-SWCNHs/S-PtNC could significantly amplify the electrochemical signal recorded by the amperometric i-t curve. Herein, a sandwich type biosensor was constructed by combining the AuNRs-SA on the electrode and c-SWCNHs/S-PtNC (signal amplifier). The label-free biosensor possessed a linear range from 5 ng mL −1 to 5 μg mL −1 with a detection limit of 0.50 ng mL −1 , and the sandwich-type biosensor possessed a wide linear range from 1 fg mL −1 to 100 ng mL −1 with a detection limit of 0.69 fg mL −1 . Furthermore, the biosensor exhibited excellent recovery and stability, indicating its potential for use in actual samples.
研究领域:
生物化学
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分析化学
·
纳米技术
·
生物传感器
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DOI
PubMed
高影响因子
ENZYME AND MICROBIAL TECHNOLOGY
·
2024
·
IF 3.4
Caixia Pei,Xinlv Peng,Yiran Wu,Runmiao Jiao,Tiehai Li,Siming Jiao,Lei Zhou,Jianjun Li,Yuguang Du,Eika W. Qian
Eukaryotic sialyltransferases play key roles in many physiological and pathological events. The expression of active human recombinant sialyltransferases in bacteria is still challenging. In the current study, the genes encoding human N -acetylgalactosaminide α2,6-sialyltransferase V (hST6GalNAc V) and N -acetylgalactosaminide α2,6-sialyltransferase VI (hST6GalNAc VI) lacking the N -terminal transmembrane domains were cloned into the expression vectors, pET-32a and pET-22b, respectively. Soluble and active forms of recombinant hST6GalNAc V and hST6GalNAc VI when coexpressed with the chaperone plasmid pGro7 were successfully achieved in Escherichia coli . Further, lactose (Lac), Lacto- N -triose II (LNT II), lacto- N -tetraose (LNT), and sialyllacto- N -tetraose a (LSTa) were used as acceptor substrates to investigate their activities and substrate specificities. Unexpectedly, both can transfer sialic acid onto all those substrates. Compared with hST6GalNAc V expressed in the mammalian cells, the recombinant two α2,6-sialyltransferases in bacteria displayed flexible substrate specificities and lower enzymatic efficiency. In addition, an important human milk oligosaccharide disialyllacto- N -tetraose (DSLNT) can be synthesized by both human α2,6-sialyltransferases expressed in E. coli using LSTa as an acceptor substrate. To the best of our knowledge, these two active human α2,6-sialyltransferases enzymes were expressed in bacteria for the first time. They showed a high potential to be applied in biotechnology and investigating the molecular mechanisms of biological and pathological interactions related to sialylated glycoconjugates.
研究领域:
分子生物学
·
生物化学
·
生物技术
·
酶学
·
糖生物学
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DOI
PubMed
高影响因子
RSC Advances
·
2016
·Vol. 6
·期 50
·44865-44872
·
IF 3.289
Yazhen Niu,Junlin He,Yuliang Li,Yilin Zhao,Chunyong Xia,Guolin Yuan,Lei Zhang,Yuchan Zhang,Chao Yu
Sialylated glycans are crucial molecular targets for cancer diagnosis and clinical research. α2,3-Sialylated glycans and α2,6-sialylated glycans are the predominant sialic acids found in nature. Different expression of the quantity of glycans can result in development of different disease. However, there are no ideal methods for discriminating α2,3-sialylated glycans and α2,6-sialylated glycans. In this work, a multi-purpose biosensor is fabricated for sensitive detection of α2,3-sialylated glycans and α2,6-sialylated glycans. To improve the sensitivity of the biosensor, p-MWCNTs were integrated with PAMAM, as PAMAM has highly branched and abundant amino groups, providing a large available surface area for linking with other substances. To achieve distinguishable recognition, Maackia amurensis lectin (MAL) and Sambucus nigra agglutinin (SNA) were included. To facilitate the lectin fixation, PDITC, a kind of green homobifunctional cross-linker, was selected. Under optimized detection conditions, the linear range of detection for α2,3-sialylated glycans is 10 fg mL−1 to 50 ng mL−1 with a lower detection limit of 3 fg mL−1, and the linear range of detection for α2,6-sialylated glycans is 10 fg mL−1 to 50 ng mL−1 with a detection limit of 3 fg mL−1. This work not only provides a method for distinguishing detection of α2,3-sialylated glycans and α2,6-sialylated glycans, but also provides a reference for future clinical testing.
研究领域:
生物化学
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分析化学
·
生物医学工程
·
临床诊断
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产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）