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Microchimica Acta

Microchimica Acta

IF: 5.29
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A simple capillary isoelectric focusing method as the novel strategy for the isoelectric point measurement of exosomes and its application in disease diagnosis

Published:5 December 2024 DOI: 10.1007/s00604-024-06864-6
Kaige Yang, Wenchang Fu, Yinjie Ma, Mingyuan Wu, Xinyan Li, Yan Wang

Abstract

A?straightforward capillary isoelectric focusing?(cIEF) method is established?with a isoelectric point?(pI) range spanning from 3.5 to 7.0, exhibiting excellent linearity and repeatability, with an R2 value of 0.9937 and migration time RSDs for all standard proteins below 0.3%. Subsequently, this method was applied to model exosomes derived from cell lines and healthy human serum, and the peak attributions of exosomes were identified using DiI labeled exosomes and lysed exosomes. The reproducibility of this method in exosome detection?was also validated, as the RSDs of all pI values were less than 1%. Then, we observed a significant increase in the pI of exosomes with higher cholesterol content, irrespective of whether they originated from cell culture medium or mouse plasma. Notably, serum exosomes from healthy volunteers exhibited higher pI values compared to those from hepatocellular carcinoma patients, suggesting a potential diagnostic perspective for cancer. These findings underscore the significance of pI measurement in reflecting modifications in exosomal lipid membrane composition and their implications in biological functions mediated by exosomes.

Graphical Abstract

Substances (17)

Materials
Procduct Name CAS Molecular Formula Supplier Price
Phosphoric acid 7664-38-2 H3O4P 1387 suppliers $33.10-$5960.00
Dimethyl sulfoxide 67-68-5 C2H6OS 1348 suppliers $15.00-$120900.00
Sodium hydroxide 1310-73-2 NaOH 1212 suppliers $14.00-$16472.00
Cholesterol 57-88-5 C27H46O 925 suppliers $10.00-$67983.00
Triethylamine 121-44-8 C6H15N 920 suppliers $9.00-$2480.00
Sodium phosphate monobasic 7558-80-7 H3O4P.Na 889 suppliers $5.00-$1130.00
Methanol 67-56-1 CH4O 838 suppliers $9.00-$7620.00
Sodium Phosphate, Dibasic 7558-79-4 Na2HPO4 662 suppliers $9.00-$4630.00
Thiourea 62-56-6 CH4N2S 593 suppliers $18.00-$1095.00
Triton X-100 9002-93-1 (C2H4O)nC14H22O 536 suppliers $18.00-$610.00

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