A novel device for cell washing, lysis and efficient extraction of intracellular proteins in a continuous automatic mode
Shi Yu, Liu Xiujie, Yuedong Chen, Yulin Deng, Rongji Dai
Abstract
Shi Yu, Liu Xiujie, Yuedong Chen, Yulin Deng, Rongji Dai
Abstract
By simultaneously subjecting cells to lysis buffer and vibration of the cell suspension fluid, effective release of intracellular contents was obtained. The synergistic effect of the applied lysis buffer and vibration on cell lysis in a membrane separation device was demonstrated for suspension cell. The resulting lysate is suitable for downstream biological sample analysis and biomedical research without requiring further separation. The cell pretreatment chamber employs a membrane separation device which provided an effective area for the cell retention, washing and lysis by peristaltic pumps and solenoid valves. To control the roller rotation direction of pump, vibrations can be produced at a certain frequency on the surface of membrane in pretreatment chamber. Then the intracellular contents were pumped into the units of filtering and ultrafiltration to remove impurities (cell fragments) and separate intracellular proteins, respectively. The result exhibits the feasibility of the proposed prototype device. The working device suggests an inexpensive, easy to be fabricated and effective way for cell pretreatment and separation of intracellular proteins, which can be easily integrated with other functional components to form a multifunctional system for analysis of biological samples.
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By simultaneously subjecting cells to lysis buffer and vibration of the cell suspension fluid, effective release of intracellular contents was obtained. The synergistic effect of the applied lysis buffer and vibration on cell lysis in a membrane separation device was demonstrated for suspension cell. The resulting lysate is suitable for downstream biological sample analysis and biomedical research without requiring further separation. The cell pretreatment chamber employs a membrane separation device which provided an effective area for the cell retention, washing and lysis by peristaltic pumps and solenoid valves. To control the roller rotation direction of pump, vibrations can be produced at a certain frequency on the surface of membrane in pretreatment chamber. Then the intracellular contents were pumped into the units of filtering and ultrafiltration to remove impurities (cell fragments) and separate intracellular proteins, respectively. The result exhibits the feasibility of the proposed prototype device. The working device suggests an inexpensive, easy to be fabricated and effective way for cell pretreatment and separation of intracellular proteins, which can be easily integrated with other functional components to form a multifunctional system for analysis of biological samples.
Key concepts: Lysis, Intracellular, Cell disruption, Membrane, Peristaltic pump, Ultrafiltration (renal), Chromatography, Cell