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J Immunoassay Immunochem ; 39(3): 308-322, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29995570

RESUMO

Fe3O4/Ag core/shell nanoparticles functionalized with the free amino (NH2) functional groups (Fe3O4/Ag-NH2) were conjugated with fluorescent electron coupled dye (ECD)-antiCD34 antibody using the 1-ethyl-3-(3'-dimethyl-aminopropyl) carbodiimide (EDC) catalyst (ECD - Electron Coupled Dye or R Phycoerythrin-Texas Red is a fluorescent organic dye attached to the antibody). The characteristic fluorescence of ECD in the antibody was investigated and was used as a good indicator for estimating the percentage of the antibodies that were successfully conjugated with the nanoparticles. The conjugation efficiency was found to increase depending on the VNP:VAB ratio, where VNP and VAB are the volumes of the nanoparticle solution (concentration of 50 ppm) and the as-purchased antibody solution, respectively. The conjugation efficiency rapidly increased from approximately 18% to approximately 70% when VNP:VAB was increased from 2:1 to 100:1, and it gradually reached the saturated state at an efficiency of 95%, as the VNP:VAB was equal to 300:1. The bioactivity of the abovementioned conjugation product (denoted by Fe3O4/Ag-antiCD34) was evaluated in an experiment for the collection of stem cells from bone marrow samples.


Assuntos
Antígenos CD34/análise , Separação Celular/métodos , Óxido Ferroso-Férrico/química , Separação Imunomagnética/métodos , Nanopartículas/química , Prata/química , Células-Tronco/citologia , Antígenos CD34/imunologia , Separação Celular/instrumentação , Humanos , Separação Imunomagnética/instrumentação , Células-Tronco/imunologia
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