ACS Biomaterials Science & Engineering
·
2020
·
IF 4.152
Honglei Wang,Xue Qu,Zheng Zhang,Miao Lei,Haoqi Tan,Chunyan Bao,Shaoliang Lin,Linyong Zhu,Joachim Kohn,Changsheng Liu
The construction of a biomaterial matrix with biological properties is of great importance to developing functional materials for clinical use. However, the site-specific immobilization of growth factors to endow materials with bioactivities has been a challenge to date. Considering the wide existence of glycosylation in mammalian proteins or recombinant proteins, we establish a bioaffinity-based protein immobilization strategy (bioanchoring method) utilizing the native sugar–lectin interaction between concanavalin A (Con A) and the oligosaccharide chain on glycosylated bone morphogenetic protein-2 (GBMP-2). The interaction realizes the site-specific immobilization of GBMP-2 to a substrate modified with Con A while preserving its bioactivity in a sustained and highly efficient way, as evidenced by its enhanced ability to induce osteodifferentiation compared with that of the soluble GBMP-2. Moreover, the surface with Con A-bioanchored GBMP-2 can be reused to stimulate multiple batches of C2C12 cells to differentiate almost to the same degree. Even after 4 month storage at 4 °C in phosphate-buffered saline (PBS), the Con A-bioanchored GBMP-2 still maintains the bioactivity to stimulate the differentiation of C2C12 cells. Furthermore, the ectopic ossification test proves the in vivo bioactivity of bioanchored GBMP-2. Overall, our results demonstrate that the tag-free and site (i.e., sugar chain)-specific protein immobilization strategy represents a simple and generic alternative, which is promising to apply for other glycoprotein immobilization and application. It should be noted that although the lectin we utilized can only bind to d-mannose/d-glucose, the diversity of the lectin family assures that a specific lectin could be offered for other sugar types, thus expanding the applicable scope further.
研究领域:
生物材料
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组织工程
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生物医学工程
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蛋白质工程
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF THE ELECTROCHEMICAL SOCIETY
·
2019
·Vol. 166
·期 2
·B49
·
IF 3.12
Di Liu,Fangxin Hu,Han Zhang,Cong Zhang,Shihong Chen
A sandwich electrochemiluminescence (ECL) biosensing system for soybean agglutinin (SBA) detection was developed with AuPt@C60 nanoflowers (AuPt@C60 NFs) as a novel co-reactant accelerator to enhance the performance of N-(aminobutyl)-N-(ethylisoluminol) (ABEI). In this strategy, nanoflower-liked AuPt@C60 complex served as a matrix for loading a large sum of ABEI and galactosamine (Gal) by Au-N and Pt-N bonds to achieve the nanocomposite Gal-ABEI-AuPt@C60 NFs. Dissolved oxygen was used as a co-reaction reagent, while AuPt@C60 complex acted as a co-reactant accelerator for dissolved oxygen to enhanced the ECL emission of ABEI. Gal, as an identifying element, was highly loaded by electrodeposited gold nanoparticles (Dp Au) on the electrode to recognize SBA, and further combine with Gal-ABEI-AuPt@C60 NFs to form a sandwich model. As a result, the proposed SBA biosensor exhibited a wide linear range from 1.0 ng·mL−1 to 5.0 μg·mL−1 and a low detection limit of 0.33 ng·mL−1. In addition, the biosensor displayed an excellent selectivity and an acceptable stability and reproducibility. The combination of AuPt@C60 NFs and ABEI provided a new ECL sensing platform without exogenous co-reactant.
研究领域:
生物传感器
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分析化学
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食品科学
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DOI
高影响因子
NANOTECHNOLOGY
·
2019
·Vol. 30
·期 48
·485706
·
IF 3.399
Feng Long,Wei Li,Wanling Chen,Danni Liu,Yue Chen,Rui Zhou,Penghui Li
In this work, a novel label-free electrochemical biosensor is developed for the detection of galectin-1 (Gal-1) based on gold nanoparticle (AuNP) loaded octahedral Cu2O (Cu2O@Au) nanocomposites. The AuNPs on the surface of the Cu2O nanocrystals not only enhance the electrochemical performance, but also serve as the binding sites for the lactose ligand which can specifically bind with Gal-1. The Cu2O@Au nanocomposites provide the synergic effect of electrochemical signal amplification and lactose–galectin reaction as the recognition strategy. Under optimal conditions, the proposed biosensor exhibits a variation of electrochemical responses to different concentrations of Gal-1 ranging from 0.1 pg ml−1 to 10 ng ml−1. This work presents an alternative electrochemical biosensor for the detection of tumor biomarkers based on a simple and economical lactose ligand incorporated Cu2O@Au biosensor platform.
研究领域:
生物传感器
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分析化学
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癌症研究
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纳米技术
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF BIOMATERIALS APPLICATIONS
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2020
·
IF 2.22
Yongli Zheng,Yina Zhang,Hua Wu,Dongjie Shao,Fawei Shen,Hao Cai,Jing Quan
Glycopolymer-grafted gold nanorods (GlycoAuNRs) were designed and fabricated as a heptoma-targeting phototherapy nanoparticles. The block glycopolymer, poly (diethyleneglycol methacrylate)-block-poly(6-O-vinyladipoyl-α-D-galactose) (PDEGMA-b-POVNGA), was synthesized by RAFT polymerization and modified AuNRs with Au-S bonds via reducing the thioester at the end of glycopolymer to thiol (–SH). Galactose pendants of thermosensitive AuNRs@PDEGMA-b-POVNGA were exposed when the temperature of aqueous dispersion is above lower critical solution temperature, which was responsible for specific binding ability with ricinus communis agglutinin. In vitro experiment indicated that the synthesized glycopolymer could significantly improve cytocompatibility of AuNRs and target to hepatocellular carcinoma cells, owing to the galactose moiety that recognized the overexpressed asialoglycoprotein receptor on the hepatocellular carcinoma cells. Interestingly, the addition of ricinus communis agglutinin could inhibit the growth of the hepatocellular carcinoma cells in some extent. Furthermore, the photothermal therapy assay proved that glycoAuNRs-ricinus communis agglutinin could realize highly effective photothermal treatment for the tumor cells in vitro upon laser irradiation by cooperative effect.
研究领域:
生物医学工程
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肿瘤学
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材料科学
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DOI
PubMed
高影响因子
FOOD AND AGRICULTURAL IMMUNOLOGY
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2012
·
IF 0.714
Xiaofang Deng,Liqiang Liu,Wenwei Ma,Chuanlai Xu,Libing Wang,Hua Kuang
In this study, two anti-peanut agglutinin (PNA) polyclonal antibodies were successfully prepared. Using mouse anti-PNA polyclonal antibody as the capture antibody and rabbit anti-PNA polyclonal antibody as the detection antibody, a novel sandwich enzyme-linked immunosorbent assay (ELISA) was developed to quantify PNA. The detection limit of the ELISA method was low (0.49 ng/mL), and the linear dynamic range was between 0.78 and 100 ng/mL. The recovery ranged from 93.86% to 139.3%, whereas the intra- and inter-assay coefficients of variation were less than 10.25% and 12.06%, respectively. Sample analysis verified this method as a reliable tool for the detection of PNA in processed foods.
研究领域:
免疫学
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食品科学
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产品关联
DOI
高影响因子
FISH & SHELLFISH IMMUNOLOGY
·
2017
·
IF 3.148
Liangliang Mu,Xiaoxue Yin,Jing Liu,Liting Wu,Xia Bian,Yuhong Wang,Jianmin Ye
Mannose-binding lectin (MBL) is a pattern recognition protein that plays an important role in innate immunity capable of activating the lectin pathway of the complement system. In this study, a MBL homologue (OnMBL) was identified from Nile tilapia ( Oreochromis niloticus ) and characterized at expression and agglutination functional levels. The open reading frame of OnMBL is 687 bp of nucleotide sequence encoding polypeptides of 228 amino acids. The deduced amino acid sequence is highly homology to teleost and similar to mammalian MBL, containing a canonical collagen-like region, a carbohydrate recognition domain and a neck region. Expression analysis revealed that the OnMBL was highly expressed in the liver, and also exhibited in other tissues including hind kidney , intestines, head kidney and spleen. In addition, the OnMBL expression was significantly up-regulated in spleen and head kidney following challenges with a Gram-positive bacterial pathogen ( Streptococcus agalactiae ) and a Gram-negative bacterial pathogen ( Aeromonas hydrophila ). Recombinant OnMBL ((r)OnMBL) protein was able to agglutinate both S. agalactiae and A. Hydrophila in vitro. Taken together, the results of this study indicated that OnMBL, possessing apparent agglutination ability to bacterial pathogens, might be involved in host defense against bacterial infection in Nile tilapia.
研究领域:
免疫学
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分子生物学
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鱼类病理学
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DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
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2019
·Vol. 411
·期 23
·6049-6056
·
IF 3.286
Zhang Cong,Hu Fangxin,Zhang Han,Chen Shihong,Yuan Ruo
As an important glycoprotein of the lectin family, soybean agglutinin (SBA) is an anti-nutritional factor with considerable toxic and side effects and plays a significant role in tumor analysis. In order to achieve the sensitive detection of SBA, a sandwich-structured electrochemiluminescence (ECL) biosensor was constructed using carboxylated carbon nitride (C-g-C 3 N 4 ) as luminophore and D-galactosamine (galM) as a recognition element. A glassy carbon electrode (GCE) was modified with Au nanoparticles (Au NPs) for capturing the galM via Au-N bond, and further capturing the target SBA by specific recognition between galM and SBA. In the presence of SBA, the composite C-g-C 3 N 4 -galM was immobilized onto the electrode. With the increase in the concentration of SBA, the ECL signal from C-g-C 3 N 4 increased, thus achieving a signal-on detection of SBA. The linear range of the biosensor was 1.0 ng/mL~10 μg/mL and detection limit for SBA was as low as 0.33 ng/mL. In this construction strategy, C-g-C 3 N 4 not only acted as an excellent signal probe, but also as an immobilization matrix to easily achieve a high loading of the small molecule recognition element galM. This strategy provides a simple alternative SBA detection platform.
Graphical abstract
研究领域:
生物传感
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食品科学
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肿瘤分析
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产品关联
DOI
PubMed
高影响因子
Chemical Science
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2011
·Vol. 2
·期 12
·2353-2360
·
IF 0
Xinai Zhang,Yingqiao Teng,Ying Fu,Shengping Zhang,Ting Wang,Chuangui Wang,Litong Jin,Wen Zhang
In this paper, we report a novel lectin-based electrochemical biosensor constructed by functionalized multiwalled carbon nanotubes (CNTs) for a competitive assay of glycan expression on living cells. The biosensor was fabricated by adsorbing poly(diallyldimethylammonium chloride) (PDDA)-functionalized CNTs (PDCNTs) onto a glassy carbon working electrode (GCE), followed by adsorbing glutathione (GSH)-protected gold nanoparticles (AuNPs). The resulting GCE/PDCNT/Au–GSH offers an effective platform for concanavalin A (Con A, a lectin) immobilization with high stability and bioactivity. A thiol-derivatized carbohydrate (thiomannosyl dimer) was synthesized to construct {CNT/thionine/Au–S–mannose} biocomposites, which exploit CNTs as carriers to load enormous amounts of thionine for generating an electrochemical signal and AuNPs for anchoring the thiomannosyl dimer. These two applications of the functionalized CNTs as a biosensing platform and biocomposites play important roles in signal enhancement for glycan detection. Using a competitive assay, the electrochemical biosensor shows good analytical performance with high sensitivity, selectivity and a rapid response for the analysis of mannose on human lung cancer cells as a model glycan. This proposed strategy possesses promising application for the assay of other glycans on living cancer cells with the selection of more lectins with the biosensor development.
研究领域:
生物化学
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医学诊断
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纳米技术
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产品关联
DOI
BIOSENSORS & BIOELECTRONICS
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2020
·
IF 10.257
Lizhen Chen,Jie Wu,Feng Yan,Huangxian Ju
This work designed a facile sensing strategy for quantitation of glycans on cell surface using organic electrochemical transistors (OECTs). The sensing strategy was performed by covalently binding target cells on mercaptopropionic acid modified gate electrode for the recognition of horseradish peroxidase (HRP) labeled lectins to specific glycans on cell surface, which led to sensitive channel current signal responding to the captured HRP and thus the expression of cell surface glycans. The quantitation of glycans on cell surface was achieved by using glycan modified microspheres to simulate the cells, on which the amounts of glycans were detected with enzyme linked immunosorbent assay . The proposed sensing strategy had been used for the detection of mannose and galactose on HeLa cells with Concanavalin A and peanut agglutinin as the specific lectins, which showed 1.37 × 10 8 mannose and 1.13 × 10 8 galactose on each cell, respectively. By treating HeLa cells with exoglycosidases , the mannose and galactose expression levels showed the same changes as those detected with flow cytometric analysis, indicating the practical application of the OECT-based biosensor in cell surface glycan expression analysis.
研究领域:
生物传感器
·
分析化学
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细胞生物学
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生物化学
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产品关联
DOI
PubMed
高影响因子
DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY
·
2018
·
IF 2.913
Liangliang Mu,Xiaoxue Yin,Yanhui Xiao,Xia Bian,Yanjian Yang,Liting Wu,Jianmin Ye
Collectins, a subfamily of the C-type lectins, are able to bind non-self glycoconjugates on the surface of microorganisms and inhibit infection by direct neutralization, agglutination and/or opsonization , which play important roles in innate immunity . In this study, a CL11X1-like collectin (OnCL11X1) was identified from Nile tilapia ( Oreochromis niloticus ) and characterized at expression and agglutination functional levels. The open reading frame of OnCL11X1 is 840 bp of nucleotide sequence encoding polypeptides of 279 amino acids . The deduced amino acid sequence is highly homology to teleost and similar to mammalian CL11X1, containing a canonical collagen-like region, a carbohydrate recognition domain and a neck region. Expression analysis revealed that the OnCL11X1 was highly expressed in the liver, and widely exhibited in other tissues including kidney, intestines and spleen. In addition, the OnCL11X1 expression was significantly up-regulated in spleen and anterior kidney following challenges with a Gram-positive bacterial pathogen ( Streptococcus agalactiae ) and a Gram-negative bacterial pathogen ( Aeromonas hydrophila ). The up-regulation of OnCL11X1 expression was also demonstrated in hepatocytes and macrophages in vitro stimulation with S. agalactiae and A. hydrophila . Recombinant OnCL11X1 protein was able to agglutinate both S. agalactiae and A. hydrophila in vitro and promote the phagocytosis by macrophages. Taken together, the results of this study indicated that OnCL11X1, possessing apparent agglutination and opsonization ability to bacterial pathogens, might be involved in host defense against bacterial infection in Nile tilapia.
研究领域:
免疫学
·
鱼类生物学
·
微生物学
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DOI
PubMed
高影响因子
COLLOIDS AND SURFACES B-BIOINTERFACES
·
2013
·
IF 3.554
Cang Wang,Yan Fan,Meng-Xin Hu,Wei Xu,Jian Wu,Peng-Fei Ren,Zhi-Kang Xu
Glycosylated membrane, as one of the most important affinity membranes, permits affinity separation/purification of proteins based on carbohydrate–protein interactions. It is an important scientific issue to screen facile method for fabricating the glycosylated membrane surface with high glycosyl density. Such a surface can be fabricated by the direct covalent immobilization of carbohydrate ligands on the surfaces of microporous polypropylene membrane (MPPM). First, alkyne-functionalized membrane surface was fabricated by plasma pretreatment combined with UV-induced graft polymerization of 3-(trimethylsilyl) propargyl methacrylate. Then, the glycosylated membrane surface was directly fabricated with the thiol–yne click reaction to ensure rapid process, improved efficiency, and high glycosyl density. Chemical and physical properties of the membrane surface were characterized by ATR/FT-IR, XPS, FESEM and water contact angle measurement. Static lectin adsorption indicates that the glycosylated membrane can specifically adsorb lectin concanavalin A (Con A) other than peanut agglutinin (PNA). Break through curves from dynamic Con A adsorption show the membrane has unique properties such as strong specificity, high adsorption capacity, and reversible binding capability. We suggest that the prepared glycosylated membrane is of great potentials in affinity membrane chromatography for rapid and high-resolution separation/purification of lectins.
研究领域:
生物化学
·
材料科学
·
分离技术
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DOI
高影响因子
ANALYTICAL CHEMISTRY
·
2015
·
IF 5.636
Xiaodong Bi,Daojin Li,Zhen Liu
Lectin array is an important tool in the fields of carbohydrate chemistry, glycobiology, and glycomics. Because natural lectins are associated with some apparent disadvantages such as tedious purification and easy loss of activity, artificial materials are applied to overcome such shortages by mimicking and replacing lectins in an artificial lectin array, among which boronate affinity-based materials are very outstanding and widely used. However, complicated synthetic works are often involved to design and create boronate affinity-based lectin-mimics. In this work, a facile and novel method was proposed to establish a virtual lectin array based on boronate affinity-based pH-featured encoding for discrimination of monosaccharides by pattern recognition. The dependence of boronate affinity on environmental pH was selected to encode each monosaccharide for feature generation, and the pH-featured encoding was used to construct the virtual lectin array. On the basis of the virtual array, pattern recognition algorithms were applied for data analysis. Monosaccharides were discriminated by principal component analysis, and the relations in the virtual lectin array were unraveled by cluster analysis. In this proof-of-concept work, without complicated synthesis or preparation, the proposed method was successful in mimicking lectin array and discriminating nine elementary monosaccharides found in nature, and it was also a new way of encoding in expanding the applications of boronate affinity-based materials and methods in the field of biomimetics.
研究领域:
糖化学
·
生物信息学
·
仿生学
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）