Isolation and quantification of human urinary exosomes by hydrophobic interaction chromatography on a polyester capillary-channeled polymer fiber stationary phase.

Abstract

Exosomes are vesicles secreted by cells having a size range from 30 to 150nm and carrying genetic materials that are important for intercellular functions, including cancer progression. Mounting evidence shows that tumor cells secrete more exosomes than normal cells. Thus, it is important to be able to efficiently isolate and quantify exosomes for potential use in clinical diagnostics, as well as to develop a deeper understanding of their role in intercellular processes. Current methods for exosome isolation and quantification are time-consuming and expensive. Few of these methods are able to combine exosome isolation and quantification into a singular operation scheme. However, a new efficient, rapid, and low-cost isolation and quantification method for exosomes in human urine samples using polyester (PET) capillary-channeled polymer (C-CP) fibers in a hydrophobic interaction chromatography (HIC) protocol has been developed. The process has been verified via scanning electron microscopy (SEM) before and after the capture of exosomes on the fiber surfaces. Sample load and elution rates were optimized to affect high resolution and throughput. Isolated exosomes were quantified based on a UV absorbance response curve created using a commercial human urine-derived exosome standard with an exosome concentration of 7.32 × 1011mL-1. The loading capacity of a 30-cm C-CP PET column was ~ 7 × 1011 exosomes. An inter-injection washing method with PBS was developed to improve the reproducibility with a 2.9% RSD achieved for 7 complete isolation cycles. Graphical abstract.