1.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
;
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