RSC Advances
·
2020
·Vol. 10
·期 47
·28041-28048
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IF 3.119
Caixia Liu,Baiping Mao,Vanessa Martinez,Xiaojian Chen,Yanan Li,Lingyun He,Sian Chen,Xiaoling Guo,Xian Shen,Xiandan Bao,Haifa Shen,Stefania Lenna,Pinyi Qian,Lingzhi Wu,Chao Li
The outbreak of new coronavirus disease (COVID-19) has quickly spread all over the world. Real time reverse transcriptase polymerase chain reaction (rRT-PCR) for nucleic acid detection has become the standard method for clinical diagnosis of COVID-19 infection. But these rRT-PCR tests have many inherent limitations, and carry a high false negative rate. It is an urgent to develop a method to accurately identify the vast infected patients and asymptomatic viral carriers from the population. In this article, we present the principle and procedure of developing a colloidal gold immunochromatographic assay (GICA) for rapid detection of COVID-19-specific antibodies. The detection kit can be used to detect immunoglobulin M (IgM) and IgG of COVID-19 in human blood samples within 15 minutes, and to identify different stages of viral infection. Test results can be digitalized using an office scanner and a FiJi software with appropriate confidence interval (CI) setting. Based on analysis from 375 samples, we calculated that overall sensitivity and specificity of the assay were 95.85% and 97.47%, respectively. Compared with rRT-PCR, this assay has many advantages including convenience and rapid detection. The detection kit can be widely used in hospitals, clinics and laboratories for rapid screening of both symptomatic and asymptomatic COVID-19 carriers in large scale.
研究领域:
医学
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病毒学
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PubMed
高影响因子
RSC Advances
·
2015
·Vol. 5
·期 82
·67093-67101
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IF 3.84
Yuping Wei,Jiandong Xu,Liang Zhang,Yankai Fu,Xia Xu
The huge demand for the most promising biopharmaceuticals, monoclonal antibodies (mAb), has resulted in a need for more efficient and less costly downstream purification processes for mAb. The currently used Protein A Agarose is unattractive due to toxic ligand leakage and high cost. In this study, three novel small peptide ligands, DWHW, CEWW and HEYW, were designed based on the interactions with human immunoglobulin G (IgG) using molecular simulations. The effects of pH, ionic strength and flow rate on the binding capacities to IgG were investigated. The static and dynamic binding capacities were determined. The dissociation constant (Kd) of the DWHW resin was 1.1 × 10−5 M. The binding capacity of the DWHW resin was 24.5 mg ml−1, which is comparable to Protein A Agarose. The DWHW resin was able to purify IgG from cMEM and CHO cell culture supernatants with a purity of more than 95% under elution conditions of pH 9 (50 mM sodium borate buffer) and pH 3 (50 mM glycine–HCl buffer). The results indicate that the small peptide ligands, especially DWHW, can offer a potential alternative for mAb purification.
研究领域:
生物化学
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生物工程
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制药工程
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高影响因子
BIOSENSORS & BIOELECTRONICS
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2021
·
IF 10.618
Quan-Quan Zhuang,Rui-Ting Chen,Yi-Jing Zheng,Kai-Yuan Huang,Hua-Ping Peng,Zhen Lin,Xing-Hua Xia,Wei Chen,Hao-Hua Deng
Assays for detecting tetanus toxoid are of great significance to be applied in the research of the safety testing of tetanus vaccine. Currently, guinea pigs or mice are usually used to evaluate the toxicity in these assays. Herein, a facile and quick biomineralization process was carried out to generate tetanus human immunoglobulin G (Tet-IgG)-functionalized Au nanoclusters (Tet-IgG-AuNCs). The obtained Tet-IgG-AuNCs exhibited strong red emission with a photoluminescence quantum yield of 13%. Based on surface plasmon resonance measurements, the apparent dissociation constant of the Tet-IgG-AuNC–tetanus toxoid complexes was measured to be 2.27 × 10 −8 M. A facile detection approach was developed using a fluorescent Tet-IgG-AuNC–based immunochromatography test strip. By utilizing the high-brightness fluorescent Tet-IgG-AuNCs, this immunosensor showed favorable sensitivity with a detection limit at the level of 0.03 μg/mL. Further results demonstrated that this assay can reliably detect tetanus toxoid and therefore might provide a novel method to replace animal tests for the quantification of tetanus toxicity. Moreover, the antibody-AuNC-based immunochromatography test strip platform serves as a promising candidate to develop new approaches for detecting targeted antigens and biological events of interest.
研究领域:
生物医学工程
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疫苗研究
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免疫学
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PubMed
高影响因子
MICROCHIMICA ACTA
·
2025
·Vol. 192
·期 7
·1-12
·
IF 5.3
Wu Han,Li Hai,Shu Jiayi,Yang Changyan,Wang Jianglong,Li Haitao,Shao Min,Zhou Hefeng
In December 2019, Wuhan, China, reported the first cases of pneumonia caused by a previously unknown coronavirus, subsequently designated SARS-CoV-2. This pathogen precipitated a global pandemic of coronavirus disease 2019 (COVID-19). The virus primarily transmits via aerosols, direct contact, and contaminated surfaces. Its high infectivity and rapid transmission have posed severe public health challenges on a global scale, emphasizing the essential need for precise and swift diagnostic techniques to effectively track and curb its transmission. To address this need, we designed a highly efficient approach, namely a colloidal gold immunochromatographic strip (CGICS) based on an IgG-mediated immunoassay, for the specific capture of the SARS-CoV-2 nucleocapsid (N) antigen. Evaluation of the assay’s specificity with a panel of common respiratory viral pathogens revealed that the CGICS specifically recognized SARS-CoV-2, with no cross-reactivity observed against non-target viruses. In addition, limit-of-detection (LOD) assessments indicated that the minimum detectable concentration was 2 ng/mL, and agreement analysis experiments showed a concordance rate of 98%, demonstrating high specificity and sensitivity. The resulting CGICS was capable of detecting SARS-CoV-2 antigen within 5–15 min. This study provides a rapid diagnostic approach for early SARS-CoV-2 infection, offering significant implications for effective disease prevention, control, and clinical diagnosis. Graphical
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TALANTA
·
2021
·
IF 6.057
Qiaoge Bayin,Lei Huang,Chunhui Ren,Yusheng Fu,Xing Ma,Jinhong Guo
Since December 2019, Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has caused millions of deaths and seriously threatened the safety of human life; indeed, this situation is worsening and many people are infected with the new coronavirus every day. Therefore, it is very important to understand patients’ degree of infection and infection history through antibody testing. Such information is useful also for the government and hospitals to formulate reasonable prevention policies and treatment plans. In this paper, we develop a lateral flow immunoassay (LFIA) method based on superparamagnetic nanoparticles (SMNPs) and a giant magnetoresistance (GMR) sensing system for the simultaneously quantitative detection of anti -SARS-CoV-2 immunoglobulin M (IgM) and G (IgG). A simple and time-effective co-precipitation method was utilized to prepare the SMNPs, which have good dispersibility and magnetic property, with an average diameter of 68 nm. The Internet of Medical Things-supported GMR could transmit medical data to a smartphone through the Bluetooth protocol, making patient information available for medical staff. The proposed GMR system, based on SMNP-supported LFIA, has an outstanding advantage in cost-effectiveness and time-efficiency, and is easy to operate. We believe that the suggested GMR based LFIA system will be very useful for medical staff to analyze and to preserve as a record of infection in COVID-19 patients.
研究领域:
医学诊断
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生物传感技术
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传染病研究
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高影响因子
Journal of Immunotoxicology
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2022
·
IF 3.439
Wenhua Zhong,Penghuan Chang,Lianfang Gan,Lifan Zhong,Zhaoxin Yang
Most current methods to assess T-cell-dependent antibody responses (TDAR) are semi-quantitative and based on measures of antibody titer generated against a standard antigen like keyhole limpet hemocyanin (KLH). The precision, sensitivity, and convenience of TDAR assays might be improved by applying rapid, sensitive, specific cytometric bead assays (CBA). In the study here, KLH antigen was covalently coupled onto the surface of cytometric beads using immune microsphere technology, and IgM antibody capture spheres were prepared for use in pretreatment processing of samples. The working parameters associated with this novel TDAR-CBA system were optimized in orthogonal experiments. The optimal concentration of the KLH coating solution in this system was 160 μg/ml, that of the anti-KLH IgG capture spheres 6.0 × 105/ml, and the optimal dilution of fluorescein isothiocyanate (FITC)-conjugated Affini-Pure Goat Anti-Mouse IgG (H + L) was 60 μg/ml. Repeated tests indicated that this approach yielded good linearity (r2 = 0.9937) method, with a within-run precision of 3.1–4.9%, and a between-run precision of 4.4–4.9%. This new approach had a limit of detection of 113.43 ng/ml (linear range = 390.63–50 000), and an interference rate of just 0.04–3.51%. Based on these findings, it seems that a new mouse TDAR assay based on CBA can be developed that would appear to be more sensitive, accurate, and precise than the current TDAR assay approaches based on traditional ELISA.
研究领域:
免疫学
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分子生物学
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分析化学
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高影响因子
ANALYTICA CHIMICA ACTA
·
2019
·
IF 5.256
Fengjuan Ding,Zhanying Chu,Quanqing Zhang,Haiyan Liu,Weibing Zhang
Hydrophilic interaction liquid chromatography is a significant strategy for the separation and enrichment of glycoproteins and glycopeptides . A layered imine-based covalent organic polymer with mesopores (denoted as p-TpBDH) was successfully fabricated by a facile solvothermal method . Then p-TpBDH-OH with abundant hydrophilic groups was evolved from p-TpBDH by the direct reduction. 36 and 40 glycopeptides were identified from IgG digests respectively by p-TpBDH and p-TpBDH-OH. Furthermore, the p-TpBDH-OH exhibits superior selectivity (IgG: BSA = 1: 250) for glycopeptides compared with the p-TpBDH. Encouragingly, a total of 463 glycopeptides assigned to 173 glycoproteins were finally identified from only 2 μL human serum by the p-TpBDH-OH. Compared with p-TpBDH, abundant hydrophilic and nitrogen-containing affinity sites of p-TpBDH-OH facilitate effective hydrophilic interaction between the polymeric material and glycopeptides. All the results demonstrate the functionalized hydrophilic covalent organic polymer has great potential in large-scale N-glycoproteomic research.
研究领域:
生物化学
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蛋白质组学
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高影响因子
JOURNAL OF CHROMATOGRAPHY A
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2016
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IF 3.926
Jia-Li Gu,Hong-Fei Tong,Dong-Qiang Lin
Magnetic particles modified with 5-amino-benzimidazole (ABI), a ligand for hydrophobic charge-induction chromatography, were prepared and used for antibody capture. In this study, with IgG as the model target, and bovine serum albumin (BSA) as the model impurity, the separation mechanism and process of IgG was investigated. The adsorption isotherms of IgG and BSA were measured, and the effects of pH were investigated in the range of pH 4.0–8.0. The maximum adsorption capacity of IgG on the particles was 180 mg/ml at pH 7.0, while low adsorption capacity of BSA (64 mg/ml) was found at pH 7.0, resulting in good selectivity. The protein-ligand interactions were elucidated by adding NaCl and glycerol. The results indicated the hydrophobic interactions were the main forces for IgG-ligand association. Moreover, the batch uptake and desorption experiments demonstrated the fast adsorption and desorption processes for IgG separation. The purity of IgG separated from mimetic serum could reach 98.6%, and the purity of monoclonal antibody (mAb) from a cell culture supernatant was 97.1%. Magnetic particles with hydrophobic charge-induction ligands showed a robust performance and could purify antibody directly from the complicated feedstock without clarification, which would improve the efficiency of antibody purification.
研究领域:
生物化学
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蛋白质纯化
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生物医学工程
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高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2023
·
IF 8.4
Fei Wang,Xin Li,Siyuan Wang,Yitao Cao,Lingqian Zhang,Yang Zhao,Xianzi Dong,Meiling Zheng,Hongyao Liu,Weier Lu,Xinchao Lu,Chengjun Huang
Integrating surface plasmon resonance (SPR) structures on fiber end facets enables the sensing system to be miniature, label-free, real-time, and remote detection. Yet, the wave vector matching condition for SPR excitation is hard to be satisfied by regulating the wave vector of light emitted from fiber ends. Here, we demonstrate a novel SPR microsensor realized by integrating a 3D micro waveguide onto the fiber tip. The mechanism of SPR excitation is illustrated by wave vector matching between the in-plane component of waveguide mode and surface plasmon polaritons . Using two-photon polymerization (2PP) and metallization process, the designed 3D waveguide probe with a length of 10 µm is fabricated on the fiber tip. Then, the microsensor was applied for the immune-sensing of anti-human IgG and human IgG interactions, and a LOD of 1.11 nM was achieved. The ultracompact structure endows with the capability for high-performance detection in the micrometre scale through a convenient “dip-and-read” manner. As a proof-of-concept for this “dip-and-read” small volume sensing, immunoassay detection in a small sample volume of 5 μL was also successfully demonstrated. The obtained results indicated that our 3D fiber-probe SPR microsensor may provide a powerful tool for small-volume biosensing applications.
研究领域:
光学
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生物传感器
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纳米技术
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分析化学
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高影响因子
ANALYTICAL CHEMISTRY
·
2014
·
IF 5.825
Rui Li,Kangbing Wu,Changxian Liu,Yin Huang,Yanying Wang,Huaifang Fang,Huijuan Zhang,Chunya Li
A novel ionic liquid, 4-amino-1-(3-mercapto-propyl)-pyridine hexafluorophosphate (AMPPH), was successfully synthesized and characterized. Subsequently, AMPPH was used as a functional monomer to fabricate AMPPH-modified gold nanoparticles (AMPPH–AuNPs) via a one-pot synthesis method. The as-prepared AMPPH–AuNPs were confirmed with transmission electron microscopy and X-ray photoelectron spectroscopy. AMPPH–AuNPs were used to construct a biocompatible interface to immobilize rabbit anti-human IgG (anti-HIgG) onto a glassy carbon electrode (GCE) surface, followed by a cross-linking step with glutaraldehyde to fabricate an anti-HIgG–AMPPH–AuNPs/GCE. The nonspecific binding sites were enclosed with bovine serum albumin (BSA) to develop an immunosensor for human IgG. Electrochemical impedance spectroscopy, cyclic voltammetry and differential pulse voltammetry were used to investigate the electrochemical properties of the developed immunosensor. The results indicate that AMPPH–AuNPs can improve the immunosensing performance. The current response of the immunosensor was found linearly related to human IgG concentration in the range of 0.1–5.0 ng mL–1 and 5.0–100.0 ng mL–1. The detection limit is estimated to be 0.08 ng mL–1 (S/N = 3). The obtained immunosensor was successfully applied to the analysis human IgG immunoglobulin in human serum, and the results were well consistent with ELISA method.
研究领域:
材料科学
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分析化学
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纳米技术
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生物传感器
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PubMed
高影响因子
TALANTA
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2021
·
IF 6.057
Ru Bai,Yue Sun,Mengyuan Zhao,Zhen Han,Juntong Zhang,Yuze Sun,Wenjing Dong,Siyu Li
Immunoglobulin G (IgG) is related to the occurrence of many diseases, such as measles and inflammatory. In this paper, IgG imprinted polymers (IgGIPs) were fabricated on the surface of nano Au/nano Ni modified Au electrode (IgGIPs/AuNCs/NiNCs/Au) via metal-free visible-light-induced atom transfer radical polymerization (MVL ATRP). The IgGIPs were prepared by IgG conjugated with fluorescein isothiocyanate (FITC-IgG) as both a template and a photocatalyst. After the templates were removed, the photocatalysts (FITC) would not remain in the polymer and avoided all the effect of catalysts on the electrode. The fabricated electrodes were examined by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), scanning electron microscope (SEM) and X-ray photoelectron spectroscopy (XPS). Under the optimized conditions, IgGIPs/AuNCs/NiNCs/Au was prepared and used as an electrochemical biosensor. The biosensor could be successfully applied for the determination of IgG by differential pulse voltammetry (DPV) measurement. The results showed that the proposed biosensor displayed a broader linear range and a lower detection limit for IgG determination when it was compared to those similar IgG sensors. The linear range from 1.0 × 10 −6 mg L −1 to 1.0 × 10 1 mg L −1 was obtained with a low detection limit (LOD) of 2.0 × 10 −8 mg L −1 ( S / N = 3). Briefly, the biosensor in this study introduced an easy and non-toxic method for IgG determination and also provided a progressive approach for designing protein imprinted polymers.
研究领域:
分析化学
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生物传感器
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聚合物科学
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电化学
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PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2017
·
IF 4.95
Haijiao Zheng,Tenggao Zhu,Xiqian Li,Jiutong Ma,Qiong Jia
A facile online method coupling polymer monolithic microextraction (PMME) with mass spectrometry (MS) was developed for the detection of Immunoglobulin G (IgG) galactosylation glycopeptides . A peanut agglutinin- β -cyclodextrin (PNA- β -CD) functionalized poly(hydroxyethyl methylacrylate-ethyleneglycol dimethacrylate) monolith was designed via a click reaction. Thanking to the specificity of PNA- β -CD for the targets, the material exhibited enhanced enrichment selectivity and extraction efficiency for IgG galactosylation glycopeptides. Under optimal conditions, the developed method gave a linear range of 0.005–5 pmol for IgG glycopeptides with the regression coefficient greater than 0.9990, and the detection limit of IgG galactosylation glycopeptides as low as 0.5 fmol was achieved. The PMME-MS method was applied to IgG galactosylation glycopeptides in real samples including human serum and acute myelogenous leukemia (AML) cell lysate. A series of unique IgG galactosylation glycopeptides were captured by the monolith in the complex samples, indicating satisfactory enrichment ability for IgG galactosylation glycopeptides. The quick and integrated online PMME-MS method exhibited high selectivity for IgG galactosylation, demonstrating its perspectives on the development and broad applications of MS in studying galactosylation proteins regulated biological processes .
研究领域:
生物化学
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分析化学
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医学
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PubMed
高影响因子
危险品化学品经营许可证（带存储）