Original scientific paper
Size Exclusion HPLC Detection of Small- size Impurities as a Complementary Means for Quality Analysis of Extracellular Vesicles
Tao Huang
; Department of Radiology and Medical Imaging, University of Virginia, Charlottesville, VA, USA
Anna B. Banisz
; Department of Radiology and Medical Imaging, University of Virginia, Charlottesville, VA, USA
Weibin Shi
; Department of Radiology and Medical Imaging, University of Virginia, Charlottesville, VA, USA
Alexander L. Klibanov
; Department of Medicine, University of Virginia, Charlottesville, VA, USA
Jiang He
; Department of Radiology and Medical Imaging, University of Virginia, Charlottesville, VA, USA
Abstract
For extracellular vesicle research, whether for biomarker discoveries or therapeutic applications, it is critical to have high-quality samples. Both microscopy and NanoSight Tracking Analysis (NTA) for size distribution have been used to detect large vesicles. However, there is currently no well-established method that is convenient for routine quality analysis of small-size impurities in vesicle samples. In this paper we report a convenient method, called ‘size-exclusion high-performance liquid chromatography’ (SE-HPLC), alongside NTA and Microscopy analysis to guide and qualify the isolation and processing of vesicles. First, the SE-HPLC analysis was used to detect impurities of small-size proteins during the ultra-centrifugation process of vesicle isolation; it was then employed to test the changes of vesicles under different pH conditions or integrity after storage. As SE-HPLC is generally accessible in most institutions, it could be used as a routine means to assist researchers in examining the integrity and quality of extracellular vesicles along with other techniques either during isolation/preparation or for further engineering and storage.
Keywords
extracellular vesicles; exosomes; HPLC; quality analysis
Hrčak ID:
161442
URI
Publication date:
1.1.2015.
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