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对量化格兰阴性细菌细胞外囊泡的方法进行比较

Chanel A Mosby¹, Natalia Perez Devia¹, Melissa K Jones¹

¹Microbiology and Cell Science Department, Institute of Food and Agricultural Sciences, University of Florida, Gainesville, FL 32611, USA.

International journal of molecular sciences

|October 28, 2023

概括

准确量化细菌细胞外囊泡是至关重要的. 微BCA试验证明了最一致的蛋白质量定量,并且与纳米粒子跟踪分析 (NTA) 相关性最好,用于估计粒子数量.

科学领域:

微生物学微生物学
生物化学生物化学
纳米技术纳米技术

背景情况:

量化细菌细胞外囊泡 (EVs) 对研究至关重要.
不一致的量化方法阻碍了跨研究的比较.
常见的方法包括脂质,蛋白质测定和纳米粒子跟踪分析 (NTA).

研究的目的:

为了比较用于量化细菌外膜囊泡的常用方法.
评估缓冲器,溶解方法和稀释剂对量化准确性的影响.
确定细菌EV量化最可靠的方法.

主要方法:

细菌外膜囊泡使用脂质 (FM 4-64),蛋白质 (microBCA,Qubit,NanoOrange) 试验和NTA.被量化.
该研究评估了缓冲器类型,缓冲器年龄,溶解方法和囊泡稀释剂的影响.
分析了蛋白质定量和NTA粒子计数之间的相关性.

主要成果:

缓冲器年龄没有显著影响囊泡量化.
溶解方法影响了Qubit和NanoOrange试验的可靠性.
微BCA测定显示了不同缓冲器和稀释剂的最小变化和高一致性.
微BCA与NTA确定的粒子数最强相关性.

结论:

微BCA试验是量化细菌EVs的可靠方法,与NTA显示一致性和强烈相关性.
在没有NTA仪器的情况下,可以使用MicroBCA来估计颗粒数量,只要适当选择缓冲器和稀释剂.
可以使用microBCA和NTA生成标准曲线,以准确量化细菌EV.

关键词:

FM 4-64 FM4-64 FM4-64 FM4-64 FM4-64 FM4-64 FM6-64 FM6-64 FM6-64 FM6-64 FM6-64 FM6-64 FM6-64 FM6-64 FM6-64 FM6-64 FM6-64 FM6-64 FM7 在NTA中,NTA是NTA.纳米色 (NanoOrange) 是一个非常小的颜色.这就是Qubit Qubit.这是一种细菌细胞外囊泡.这是一个微型BCA.纳米粒子跟踪分析分析在外膜囊泡中,外膜囊泡囊泡的量化量化方法

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