Microchip Capillary Electrophoresis: Quantum Dots and Paramagnetic Particles for Bacteria Immunoseparation

Microchip Capillary Electrophoresis: Quantum Dots and Paramagnetic Particles for Bacteria Immunoseparation

Kapitola, resp. kapitoly v odborné knize

The emergence of drug-resistant bacteria and new or changing infectious pathogens is an important public health problem as well as a serious socioeconomic concern. Immunomagnetic separation-based methods create new possibilities for rapidly recognizing many of these pathogens. Nanomaterial-based techniques including fl uorescent labeling by quantum dots as well as immunoextraction by magnetic particles are excellent tools for such purposes. Moreover, the combination with capillary electrophoresis in miniaturized microchip arrangement brings numerous benefi ts such as fast and rapid analysis, low sample consumption, very sensitive electrochemical and fl uorescent detection, portable miniaturized instrumentation, and rapid and inexpensive device fabrication. Here the use of superparamagnetic particle-based fully automated instrumentation to isolate pathogen Staphylococcus aureus and its Zn(II)-containing proteins (Zn-proteins) is reported using a robotic pipetting system speeding up the sample preparation and enabling to analyze 48 real samples within 6 h. Cell lysis and Zn-protein extractions were obtained from a minimum of 100 cells with the suffi cient yield for SDS-PAGE (several tens ng of proteins).

The emergence of drug-resistant bacteria and new or changing infectious pathogens is an important public health problem as well as a serious socioeconomic concern. Immunomagnetic separation-based methods create new possibilities for rapidly recognizing many of these pathogens. Nanomaterial-based techniques including fl uorescent labeling by quantum dots as well as immunoextraction by magnetic particles are excellent tools for such purposes. Moreover, the combination with capillary electrophoresis in miniaturized microchip arrangement brings numerous benefi ts such as fast and rapid analysis, low sample consumption, very sensitive electrochemical and fl uorescent detection, portable miniaturized instrumentation, and rapid and inexpensive device fabrication. Here the use of superparamagnetic particle-based fully automated instrumentation to isolate pathogen Staphylococcus aureus and its Zn(II)-containing proteins (Zn-proteins) is reported using a robotic pipetting system speeding up the sample preparation and enabling to analyze 48 real samples within 6 h. Cell lysis and Zn-protein extractions were obtained from a minimum of 100 cells with the suffi cient yield for SDS-PAGE (several tens ng of proteins).

Magnetic particles , Immunoseparation , Staphylococcus aureus , Zn-protein

Magnetic particles , Immunoseparation , Staphylococcus aureus , Zn-protein

KŘÍŽKOVÁ, S.; NGUYEN, H.; STANISAVLJEVIC, M.; KOPEL, P.; VACULOVIČOVÁ, M.; ADAM, V.; KIZEK, R.

2016

01.07.2015

Springer Science+Business Media New York 2015

978-1-4939-2353-3

Microchip Capillary Electrophoresis Protocols

67

79

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