1.Preparation and properties of SiO2 tubes immobilized antibody for HCAg detection.
Li XIE ; Yueping GUAN ; Ying GE ; Hongbo SHI
Chinese Journal of Biotechnology 2010;26(4):545-549
In order to optimize the fabrication of SiO2 tubes immobilized with antibody for hepatitis C virus antigen (HCAg) detection, we formed the activated amino on the surface of SiO2 tubes by using the activation of aminosilane. Then we immobilized the hepatitis C virus (HCV) monoclonal antibody on the surface of SiO2 tubes by using glutaraldehyde as a chemical cross-linker, followed by detecting HCAg. Sequence tests showed that when the SiO2 tubes were treated in 10% (V/V) aminosilane solution and 3% (V/V) glutaraldehyde solution for 3 hours and 2 hours, respectively, the HCV monoclonal antibody had high immobilization efficiency and low nonspecificity, and the HCAg was detected to 1 ng/mL. This experiment can provide principle and experimental data for establishment of HCAg magnetic immunoassay system.
Antibodies, Immobilized
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immunology
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Antibodies, Monoclonal
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chemistry
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immunology
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Hepatitis C Antibodies
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chemistry
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immunology
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Hepatitis C Antigens
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analysis
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immunology
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Humans
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Silicon Dioxide
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chemistry
2.Construction of gold film solid-phase carrier of myoglobin monoclonal antibody based on thiol self-assembly.
Bo LEI ; Bingjun QU ; Hua YANG ; Xiangying ZHANG ; Fang LIU
Chinese Journal of Biotechnology 2011;27(10):1527-1535
In order to immobilize myoglobin (Mb) monoclonal antibody on gold film solid-phase carrier, we grew a mixed self-assembled monolayers (SAMs) of acid thiol and mercapto ethanol on gold film. Then we analyzed the property of the sample by atomic force microscopy and X-ray photoelectron spectroscopy. Then, we used 1-(3-dimethyl aminopropyl)-3-ethyl carbodiimide hydrochloride as catalyst to couple SAMs with amino of antibody so that we immobilized antibody on surface of gold film, followed by detecting myoglobin antigen. Results showed that, by optimizing experimental conditions, when we treated gold film by a mixture of mercapto hexadecanoic acid and mercapto undecanol ethanol solution of concentration of 50 mmol/L at temperature of 60 degrees C for 3 hours, and Mb monoclonal antibody of concentration of 40 mg/L for 3 hours, respectively, antibody had high immobilization efficiency and the MbAg was detected to 30 microg/L. The method provided a theoretical and practical basis for using magnetoresistence biosensors to diagnosis myocardial infarction.
Antibodies, Immobilized
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Antibodies, Monoclonal
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immunology
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Biosensing Techniques
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instrumentation
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methods
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Gold
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chemistry
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Humans
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Membranes, Artificial
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Microscopy, Atomic Force
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Myocardial Infarction
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diagnosis
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Myoglobin
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blood
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immunology
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Photoelectron Spectroscopy
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Sulfhydryl Compounds
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chemistry
3.Serotype- and serogroup-specific detection of African horsesickness virus using phage displayed chicken scFvs for indirect double antibody sandwich ELISAs.
Wouter VAN WYNGAARDT ; Cordelia MASHAU ; Isabel WRIGHT ; Jeanni FEHRSEN
Journal of Veterinary Science 2013;14(1):95-98
There is an ongoing need for standardized, easily renewable immunoreagents for detecting African horsesickness virus (AHSV). Two phage displayed single-chain variable fragment (scFv) antibodies, selected from a semi-synthetic chicken antibody library, were used to develop double antibody sandwich enzyme-linked immunosorbent assays (DAS-ELISAs) to detect AHSV. In the DAS-ELISAs, the scFv previously selected with directly immobilized AHSV-3 functioned as a serotype-specific reagent that recognized only AHSV-3. In contrast, the one selected with AHSV-8 captured by IgG against AHSV-3 recognized all nine AHSV serotypes but not the Bryanston strain of equine encephalosis virus. Serving as evidence for its serogroup-specificity. These two scFvs can help to rapidly confirm the presence of AHSV while additional serotype-specific scFvs may simplify AHSV serotyping.
African horse sickness virus/*isolation & purification
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Animals
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Antibodies, Immobilized
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Antibodies, Viral/*immunology
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Cercopithecus aethiops
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Chickens
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Enzyme-Linked Immunosorbent Assay/methods/*veterinary
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Immunoglobulin G
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*Peptide Library
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Serologic Tests/methods/veterinary
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Serotyping
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Single-Chain Antibodies/*immunology
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Vero Cells
4.Immobilization of CD34 antibodies onto ethylene-vinyl acetate copolymer coatings and in vitro cell attachments.
Min YIN ; Yuan YUAN ; Changsheng LIU ; Jing WANG
Journal of Biomedical Engineering 2008;25(5):1130-1134
Rapid re-endothelialization of implanted coronary artery stent is a new way for preventing restenosis. In this paper, adhesive polypeptide mimics of marine organism and antibodies potentially selective targeting with CD34 antigens on endothelial cells (ECs) were immobilized onto ethylene-vinyl acetate copolymer (EVA) coatings for improving the re-endothelialization process. The results showed that the attachments, growths, viabilities, as well as cell retentions of ECs on EVA were improved after modification. The immobilized polypeptide promoted the attachments of ECs, but overmuch polypeptide would restrict the attachments. With the increase of antibody concentration, the immobilized antibodies and cell attachments were improved.
Antibodies, Immobilized
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immunology
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Antigens, CD34
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immunology
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Cell Adhesion
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drug effects
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Cells, Cultured
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Coated Materials, Biocompatible
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chemistry
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Coronary Restenosis
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prevention & control
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Endothelial Cells
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cytology
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Humans
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Peptides
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chemistry
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Polyvinyls
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chemistry
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Stents
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Umbilical Veins
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cytology