| • 0.00 min | Inject sample, 100% buffer A, 0.5 ml min−1 |
| • 6.30 min | Start to collect flow-through fraction (low-abundance proteins): 100% Buffer A, 0.5 ml min−1 |
| • 25.0 min | 100% Buffer A, 0.5 ml min−1 |
| • 25.1 min | 100% Buffer B, 0.5 ml min−1 |
| • 26.3 min | Stop to collect flow-through fraction: 100% buffer B, 0.5 ml min−1 |
| • 27.0 min | 100% Buffer B, 3.0 ml min−1 |
| • 29.0 min | Start to collect bound fraction (high-abundance proteins): 100% Buffer B, 3.0 ml min−1 |
| • 33.0 min | Stop to collect bound fraction: 100% buffer B, 3.0 ml min−1 |
| • 35.0 min | 100% Buffer B, 3.0 ml min−1 |
| • 35.1 min | 100% Buffer A, 3.0 ml min−1 |
| • 45.0 min | 100% Buffer A, 3.0 ml min−1 |