.png){kind=icon}
.png){kind=icon}
Return
A Novel Fully Automatic Free-Flow Isoelectric Focusing System and Its Application in the Large-Scale Isolation of Milk Exosomes
.png){kind=icon}
PRE
AI
Y
S
Q
X
DOI：10.1002/elps.70127.png)
.png){kind=icon}
Original request for help.png){kind=icon}
Share.png){kind=icon}
Save
Abstract
En 中文
Free-flow electrophoresis is a core technology for biological particle separation, but traditional devices suffer from low automation, poor flow stability, and limited throughput, hindering industrial-scale applications. Herein, a novel fully automatic reciprocating free-flow isoelectric focusing (RFFIEF) system, a batch-type of automated closed-loop free-flow isoelectric focusing (FFIEF), was developed, integrating a 24-channel separation chamber, closed gas–liquid circulation drive, and intelligent control system to achieve unattended standardized operation. Standard protein verification showed excellent flow stability and pH gradient reproducibility (pooled standard deviation [SD] = 0.19 pH units excluding ampholyte-induced outliers). The system was applied to large-scale separation of milk exosomes, which were effectively focused at pH 5.0–7.5. Exosome subpopulations were fractionated by surface charge: CD81-positive subpopulations concentrated at pH 5.8–6.5, whereas CD63-positive subpopulations were highly enriched at pH 6.9. This system breaks through the automation bottleneck of conventional devices, providing an efficient platform for large-scale biological particle separation with broad industrial prospects.
Keywords:
automatic control
biological particle separation
free-flow electrophoresis
fully automatic device
reciprocating isoelectric focusing
Journal
IF:
2.5
Papers:
1.2W
Citations:
1.0W
Organization
Cited Papers