ANALYTICAL BIOCHEMISTRY
·
2018
·
IF 2.275
Lide Gu,Wanli Yan,Shu Liu,Wei Ren,Mingsheng Lyu,Shujun Wang
A novel screening method for protein aptamer selection was developed in this study. Aptamers with high affinity and specificity to the surface recombinant antigen of Helicobacter pylori (HP-Ag) and to tumor markers carcinoembryonic antigen (CEA), cancer antigen 125 (CA125) and cancer antigen 19-9(CA19-9) were screened using trypsin enhanced screening method. Briefly, the target proteins above were immobilized onto 96-well polystyrene plates and incubated with a single-stranded DNA (ssDNA) library for aptamer selection. Then, trypsin was introduced to digest the proteins and obtain ssDNA that bound to the target proteins with high specificity. The concentration of ssDNA that shed from protein-ssDNA complexes was detected. After sequencing, the enrichment of target-specific aptamers was monitored and the affinity of each aptamer was analyzed. Urea, which has been reported in other article, was used to compare with trypsin. The results revealed that trypsin was more effective than urea for protein aptamer selection. The protocol used in this study provided a novel method for generating aptamers.
研究领域:
分子生物学
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生物技术
·
蛋白质工程
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DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2017
·Vol. 184
·期 11
·4269-4277
·
IF 4.58
Zheng Yun,Wang Huiqiang,Ma Zhanfang
The authors describe a method for signal amplification of label-free voltammetric immunosensors. A glassy carbon electrode (GCE) was modified with Prussian Blue-platinum nanoparticles (PB-PtNPs) as a redox-active species that gives a strong amperometric signal at 0.18 V (vs. Ag/AgCl). Benefitting from the excellent electrical conductivity and the strong catalytic activity to H 2 O 2 , the modified GCE gives a strongly enhanced signal. The PB-PtNPs were incorporated into a polyaniline (PANI) hydrogel to further enhance the signal. The signal response of the PB-PtNP-PANI/GCE is larger by a factor of 7.6 than that of PB-PtNP/GCE. In order to further improve electrical conductivity and immobilize antibody, gold nanoparticles (AuNPs) were deposited on the surface of the PB-PtNP-PANI hydrogel. The AuNP-PB-PtNP-PANI hydrogel nanocomposite on the GCE was used in an immunosensor for the model analyte carcinoma antigen 125 (CA125), a biomarker for epithelial ovarian cancer, by immobilizing the respective antibody on the modified GCE. A linear response found for the 0.01 to 5000 U mL −1 CA125 concentration range, with a detection limit of 4.4 mU mL −1 (at an S/N ratio of 3). The electrochemical sensitivity is as high as 119.76 μA·(U/mL) −1 ·cm −2 . The detection of CA125 in human serum showed satisfactory accuracy compared to a commercial chemiluminescent microparticle immunoassay (CMIA).
Schematic of a nanocomposites consisting of gold nanoparticles, Prussian Blue, platinum nanoparticles and polyaniline hydrogel as a signal multi-amplification sensing substrate for the ultrasensitive immuno detection of carcinoma antigen 125 (CA125).
研究领域:
生物医学工程
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电化学传感
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肿瘤标志物检测
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DOI
高影响因子
MICROCHEMICAL JOURNAL
·
2025
·
IF 5.1
Zhi Luo,Jing Li,Shenglan Hu,Lingling Zheng,Lixin Xu,Biyang Deng
In this study, Ru(bpy) 3 2+ and oxalate (C 2 O 4 2− ) in Ru@Zn-MOF generated self-enhanced electrochemiluminescence (ECL). The self-enhanced ECL composite exhibited a short electron transfer distance and minimal energy loss, which ensured the high and stable luminescence efficiency of Ru@Zn-MOF. A MnO 2 @AuNPs nanocomposite was introduced into Ru@Zn-MOF via antigen–antibody interactions in the presence of squamous cell carcinoma antigen (SCCA). This significantly quenched the ECL signal due to the combined effect of the dual quenchers, MnO 2 NFs and AuNPs. A highly efficient ECL-RET immunosensor was constructed for the sensitive detection of SCCA in the linear range of 1 fg/mL to 100 ng/mL, with a limit of detection of 0.26 fg/mL (S/N = 3). This immunosensor has broad clinical applications for the quantitative detection of biomarkers.
研究领域:
生物医学工程
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分析化学
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肿瘤学
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DOI
高影响因子
ANALYTICA CHIMICA ACTA
·
2020
·
IF 5.977
Ruidi Shen,Ji Zhang,Wenxiu Huang,Shaoyong Wu,Gongke Li,Seyin Zou,Liansheng Ling
Cancer antigen 125 (CA - 125) is an important biomarker for the diagnosis of ovarian cancer. In this paper, oligonucleotide 5′-GACAGGCCCGAAGGAATAGATAATACGACTCACTATAGGGAGACAAGAATAAACGCTCAA-3′ (oligo 1) contains an aptamer of CA - 125, and was designed partly complementary to oligonucleotide 5′-CTCTCTCTCCACCTTCTTCTTTGAGCGTTTATTCTTGTCT-3′ (oligo 2). Oligo 1 · oligo 2 was extended with the Klenow fragment (exo − ) polymerase for further polymerase chain reaction (PCR) processes in the presence of two primers: deoxyribose nucleoside triphosphate and Taq polymerase . Single-stranded DNA was produced at two sides of the PCR product by introducing a C 18 spacer into the two primers, which could hybridize with AuNPs-DNA probes, investigated by dynamic light scattering and fluorescence. The addition of CA - 125 can interrupt the hybridization between oligo 1 and oligo 2, causing the average diameter of AuNPs-DNA probes to decrease with the increase of CA-125 within the range of 5 fg mL −1 – 50 ng mL −1 . The linear regression equation of this relationship was D = 430.48–49.60 log 10 C, with a detection limit of 1.1 fg mL −1 . Fluorescein molecules were modified at the end of the forward primer. The fluorescence intensity of the PCR product can be measured simultaneously, with the fluorescence intensity increasing linearly with the logarithm of CA-125 concentration within a linear range from 10 fg mL −1 to 50 ng mL −1 , with a detection limit of 1.5 fg mL −1 .
研究领域:
生物化学
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分子生物学
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医学诊断
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DOI
PubMed
高影响因子
RSC Advances
·
2016
·Vol. 6
·期 9
·6932-6938
·
IF 3.289
Yueyun Li,Yihe Zhang,Liping Jiang,Paul K. Chu,Ynhui Dong,Ping Wang
An ultrasensitive electrochemical immunosensor is designed and constructed for the quantitative detection of squamous cell carcinoma antigens (SCCAs). Silver nanoparticles (AgNPs) hybridized and 3-aminopropyltriethoxysilane functionalized graphene sheets (Ag@APTES-GS) serve as the platform to immobilize primary antibodies (Ab1). The mesoporous AgNPs-decorated silica KIT-6 (Ag@KIT-6) with a large specific surface area is utilized to label the electron mediator thionin (TH) and secondary antibodies (Ab2) by physical adsorption and chemical bonding. Carboxymethyl chitosan (CMC) doped with ionic liquids (ILs) is used to facilitate electron transfer and prevent leaking of TH resulting in multiple signal amplification. In addition to the excellent detection limit (17 fg mL−1, S/N = 3) and wide linear range (5 × 10−5 ng mL−1 to 102 ng mL−1), the immunosensor shows good selectivity, reproducibility, and stability. Providing quantitative detection of SCCA in serum, the sensor has great potential in clinical and diagnostic applications.
研究领域:
生物医学工程
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诊断学
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纳米技术
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DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2016
·
IF 4.758
Shenguang Ge,Linlin Liang,Feifei Lan,Yan Zhang,Yanhu Wang,Mei Yan,Jinghua Yu
The hybrids, N -aminobutyl- N -ethylisoluminol (ABEI), horseradish peroxidase (HRP) and cancer antigen 125 s antibody (Ab2) immobilized graphene oxide (GO), were prepared, which exhibited excellent chemiluminescence (CL) activity. The zinc oxide nanorods (ZNRs) grew on reduced graphene oxide (RGO) modified paper working electrode (ZNRs-RGO/PWE) via an in situ growth method. The deposition of CdS quantum dots (QDs) on ZNRs/RGO-PWE resulted in an enhanced excitation and photo-to-electric conversion efficiency. The ABEI-GO@HRP/Ab2CL system was employed as an internal excited light source. A photoelectrochemical (PEC) immunosensor based on CdS-sensitized ZNRs anchored RGO as the photoactive matrix and ABEI-GO@HRP/Ab2 as the CL labels was established for sensitive detection of cancer antigen 125 (CA 125). As a model, sandwich CA 125-antibody was taken as molecular reorganization elements on this sensor for the sensitive determination of CA 125 in the linear range from 5.0 × 10 −4 U/mL to 500 U/mL with a detection limit of 2.0 × 10 −4 U/mL. The specificity, reproducibility, and stability of this sensor were also investigated. This proposed method might be attractive for clinical and biomedical applications.
研究领域:
生物医学工程
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分析化学
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DOI
高影响因子
BIOSENSORS & BIOELECTRONICS
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2015
·
IF 6.409
Yuanyuan Liu,Hongmin Ma,Jian Gao,Dan Wu,Xiang Ren,Tao Yan,Xuehui Pang,Qin Wei
In this study, a novel and ultrasensitive sandwich-type electrochemical immunosensor was designed for the quantitative detection of squamous cell carcinoma antigen (SCCA) based on the β-cyclodextrin functionalized graphene nanosheet (CD-GN) and the ternary hollow Pt/PdCu nanocube anchored on three-dimensional graphene framework (Pt/PdCu-3DGF). CD-GN exhibited high specific surface area and good dispersibility and stability in water, which were beneficial to fix captured antibodies (Ab 1 ) through the supramolecular host-guest interaction between CD and Ab 1 . The abundant oxygen-containing functional groups on 3DGF provided binding sites for anchoring noble metal nanoparticles . Pt/PdCu-3DGF could capture detected antibodies via the interaction of Pd-NH 2 and Pt-NH 2 . Furthermore, the ternary metal nanoparticles exhibited high electrocatalytic activity toward the reduction of hydrogen peroxide. Under optimal conditions, the fabricated immunosensor showed a sensitive response to SCCA with two linear ranges. The linear ranges are 0.0001–1 ng/mL and 1–30 ng/mL with a detection limit of 25 fg/mL. Additionally, the proposed immunosensor showed good reproducibility and stability.
研究领域:
生物化学
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医学检测
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纳米技术
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产品关联
DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
·
2024
·
IF 4.9
Fangfang Wu,Hongmin Gao,Ren Qiu,Hehua Zhang,Xinshui Ren,Xue Qi,Men Miao,Chuang Rui,Dong Chang,Hongzhi Pan
Accurate and sensitive detection of cancer antigen 125 (CA125) is important in early ovarian cancer diagnosis and monitoring. In the present work, a novel electrochemical ratiometric immunosensor for the determination of CA125 based on three-dimensional reduced graphene oxide/multi-walled carbon nanotube carboxylic acid- thionine (3DrGO/MWCNTs-Thi) and UIO-66-NH 2 /ferrocenecarboxylic acid (UiO-66-Fc) was constructed. 3DrGO/MWCNTs-Thi of immobilized primary antibody (Ab 1 ) and UiO-66-Fc of labeled second antibody (Ab 2 ) were used as capture probes and signaling probes for CA125, respectively. As the concentration of CA125 increases, the oxidation peak currents (I Thi and I Fc ) generated by the two materials were changed in opposite directions, and therefore the ratio of the currents (I Thi /I Fc ) was negatively correlated with the concentration of CA125. Thanks to the amplification of high loading of signaling molecules by the framework nanocomposites and the ratiometric strategy, the constructed electrochemical immunosensor exhibits outstanding accuracy, stability, and sensitivity. Under optimal conditions, the detection range spanned from 0.01 U mL −1 to 80 U mL −1 , with a calculated limit of detection (LOD) of 0.0089 U mL −1 . The immunosensor was successfully applied to real serum sample tests, indicating its potential for clinical detection applications.
研究领域:
生物医学工程
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分析化学
·
肿瘤学
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DOI
高影响因子
MICROCHIMICA ACTA
·
2017
·Vol. 184
·期 9
·3565-3572
·
IF 4.58
Li Mingshi,Wang Yanying,Ye Xiaoxue,Wang Zhengguo,Wu Tsunghsueh,Li Chunya
Icosahedral gold nanocrystals (AuNCs) were synthesized by using the ionic liquid (IL) 1-(10-bromodecyl)-3-methyl-imidazolium bromide as a morphology-controlling reagent. The novel IL 1,3-di-(3-mercaptopropyl)-imidazolium bromide was employed as a bifunctional monomer to integrate the AuNCs onto the surface of a glassy carbon electrode (GCE) modified with gold nanoparticles. After interaction with thioglycolic acid (TGA) and activation of the carboxy groups, the antibody against squamous cell carcinoma antigen was immobilized on the modified GCE. Residual active sites were blocked with bovine serum albumin. The new materials and the steps for immunosensor fabrication were fully characterized. The reduction in peak current (using hexacyanoferrate as an electrochemical probe) due to the recognition of SCCA is linearly related to the SCCA concentration in the 0.02 to 10 ng mL −1 concentration range. The detection limit is as low as 12.6 pg mL −1 (at an S/N ratio of 3). The assay was applied to the quantitation of SCCA in human serum samples. The accuracy was validated by comparison with an ELISA method. The two methods displayed excellent consistency.
Icosahedral gold nanocrystals were self-assembled onto a gold nanoparticle modified glassy carbon electrode surface by using 1,3-di-(3-mercaptopropyl)-imidazolium bromide ionic liquid as a linker. Antibody for squamous cell carcinoma antigen was immobilized onto the gold nanocrystal based interface to fabricate an immunosensor.
研究领域:
材料科学
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生物医学工程
·
分析化学
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DOI
高影响因子
JOURNAL OF THE ELECTROCHEMICAL SOCIETY
·
2024
·Vol. 171
·期 3
·037526
·
IF 3.1
Deyong Kang,Shanbao Ma,Qiaoling Zheng
Squamous cell carcinoma antigen (SCCA) is an important biomarker in the diagnose of cancer, so developing effective methods for its detection is of great significance. In the present work, a novel sandwich-like electrochemical immunosensing assay (STEM) of SCCA was constructed by preparing gold nanoparticle/reduced graphene-oxide (Au NPs/rGO) to immobilize primary antibody (PAb) of SCCA and UIO-66-NH2 MOFs structure to immobilize second antibody (SAb) and electroactive toluidine blue (Tb) probe. In this designed STEM assay, the prepared Au NPs/rGO nanohybrid offers the advantages in large surface area and conductivity as carrier, and UIO-66-NH2 provided an ideal platform to accommodate SAb and a large number of Tb molecules as signal amplifier. In the existence of SCCA, the current peaks of Tb from the formed STEM structure increase with the increase of SCCA level. After optimizing the related control factors, a wide linear range (0.01 pg ml−1 and 15.0 ng ml−1) and very low detection limit (0.003 pg ml−1) of SCCA was achieved, it's thus expected the designed STEM strategy has important applications for the detection of SCCA.
研究领域:
生物医学工程
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肿瘤学
·
材料科学
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产品关联
DOI
高影响因子
TALANTA
·
2014
·
IF 3.511
Yulan Wang,Yong Zhang,Yu Su,Feng Li,Hongmin Ma,He Li,Bin Du,Qin Wei
Determination of squamous cell carcinoma antigen (SCCA) in human serum plays an important role in diagnosis of cervical squamous cell carcinoma. In this work, Au/Ag/Au core/double shell nanoparticles (Au/Ag/Au NPs) were prepared by a simple approach and were used as novel enzyme-mimetic labels for development of a sandwich-type electrochemical immunosensor for SCCA. The nanostructure of Au/Ag/Au NPs could be well confirmed by transmission electron microscope (TEM) and UV–vis spectra. Au NPs decorated mercapto-functionalized graphene sheets (Au@SH-GS) were used as platform for immobilization of primary antibody (Ab 1 ), while Au/Ag/Au NPs were employed as labels of secondary antibody (Ab 2 ). Due to the excellent electrocatalytic activity of Au/Ag/Au NPs towards the reduction of hydrogen peroxide (H 2 O 2 ), electrochemical amperometric responses to SCCA were achieved after the immuno-reaction. Under optimum conditions, the electrochemical immunosensor exhibited a wide linear range from 0.5 pg/mL to 40 ng/mL with a low detection limit of 0.18 pg/mL for SCCA. The designed immunosensor displayed an excellent analytical performance with good reproducibility, high selectivity and stability.
研究领域:
生物医学工程
·
分析化学
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2018
·
IF 5.667
Yingyi Wang,Shanshan Wang,Chunsong Lu,Xiaoming Yang
Malignant tumors posed severe threat to human health, and various efforts have been contributed towards assaying tumor markers, howbeit, achieving to identify the multiple targets by one single analytical mode is in demand. Hereby, three kinds of aptamer-functionalized silver and silver/gold nanoclusters (DNA-AgNCs and DNA-Ag/AuNCs) were originally synthesized, and possessed the specific recognition ability for their corresponding targets. Specifically, the fluorescence of AgNCs or Ag/AuNCs was quenched by introducing graphene oxide (GO), and more competitive interactions between tumor markers and their aptamers occurred with the targets added, thus facilitating their fluorescence recovered. Based on AgNCs or Ag/AuNCs cooperating with GO, this sensing mode was subsequently employed for detecting mucin 1 in a linear range of 1.33–200 ng/mL with a detection limit of 0.18 ng/mL. Meanwhile, this method was also applied to examine carcinoembryonic antigen and cancer antigen 125, and showed the linear range of 6.7 ng/mL–13.3 μg/mL with a detection limit of 3.18 ng/mL for CEA, while CA125 sensing acquired a linear range of 2 ng/mL–6.7 μg/mL accompanied by a detection limit of 1.26 ng/mL. Besides, this strategy provided the potential ways to identify other tumor markers by designing their corresponding nanoclusters with the aptamers.
研究领域:
纳米技术
·
生物医学工程
·
分析化学
·
癌症研究
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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）