Table 1.
Comparison of Precipitation-Based Protocols with Basic Protocol
| protein (pmol/mg total protein)a | ||||||
|---|---|---|---|---|---|---|
|
|
||||||
| protein | Exospin (protocol #1) | Exospin (protocol #2) | total Exo (protocol #3) | total Exo (protocol #4) | EVs (208 K) | EVs (SEC) |
| albumin | 3163.0 ± 817.5 | 1592.0 ± 284.8 | 1119.0 ± 1.4 | 603.0 ± 222.5 | 6839.0 ± 177.4 | 3316.0 ± 40.3 |
| transferrin | 109.6 ± 15.3 | 22.9 ± 1.9 | 25.4 ± 4.9 | 32.8 ± 5.7 | 154.2 ± 0.01 | 35.1 ± 0.1 |
| α-2-macroglobulin | 4038.0 ± 254.5 | 6529.0 ± 504.2 | 1132.0 ± 56.6 | 676.0 ± 119.3 | 270.0 ± 18.0 | 133.5 ± 7.9 |
| apoE | 79.2 ± 5.8 | 121.9 ± 7.4 | 81.8 ± 7.0 | 480.9 ± 28.9 | 102.8 ± 0.03 | 314.0 ± 0.1 |
| HSPA8 | 0.12 ± 0.05 | 0.11 ± 0.03 | 0.08 ± 0.02 | 0.09 ± 0.05 | 0.50 ± 0.02 | 6.39 ± 2.4 |
| moesin | 0.67 ± 0.20 | 0.76 ± 0.16 | 0.49 ± 0.17 | 0.54 ± 0.31 | 2.59 ± 0.12 | 4.34 ± 0.10 |
| HSP90AA1 | 0.05 ± 0.07 | 1.53 ± 0.05 | ||||
| EHD4 | 0.03 ± 0.04 | 0.08 ± 0.01 | ||||
| flotillin-1 | 0.02 ± 0.05 | 0.72 ± 0.02 | ||||
| HDAC 1,2 | 0.06 ± 0.03 | 0.38 ± 0.03 | ||||
| HSP90AB1 | 4.66 ± 0.11 | |||||
| TSG101 | 0.39 ± 0.01 | |||||
| DNAJB1 | 0.25 ± 0.20 | |||||
| L1CAM | 0.25 ± 0.13 | |||||
a
MRM measurements were normalized to the total protein in each sample. Exospin Blood kit was from Cell Guidance Systems and total exosome isolation (from serum) kit was from Invitrogen. Protocols #1 through #4 are summarized in Figure S3 (Supporting Information). All experiments were performed in duplicate with three replicate injections to assess error and reproducibility. Data is represented as the mean ± SD.