SAX-ERLICH Enrichment

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Strong anion exchange chromatography using ERLIC (Totten et al., 2017) (SAX-ERLIC) was performed on two aliquots following immunodepletion, with no prior desalting with C18 SPE. SOLA SAX SPE cartridges (ThermoFisher Scientific) were loaded on a vacuum manifold. The cartridges were preconditioned using in-house vacuum during washing. One mL of each wash solution was applied at a time including 3 mL of ACN, followed by 3 mL of 100 mM triethylammonium acetate in water, 3 mL of 1% TFA in water, and 3 mL of 95% ACN with 1% TFA in water. The organic content of the tryptic peptide solution was adjusted to 95% ACN with 1% TFA in water. Peptides were applied to the SPE cartridge; then, the cartridge was washed with 500 μL of 95% ACN with 1% TFA in water. Positive pressure from a pipet bulb was used to assist with flow across the SPE cartridge. The SPE cartridge was then washed with an additional 6 mL of 95% ACN with 1% TFA in water. The peptides were next eluted in three 500 μL aliquots of 50% ACN with 0.1% TFA in water. Larger peptides were eluted in two 1 mL aliquots of 5% ACN with 0.1% TFA in water. Eluates were combined and dried in a Speedvac.

References

Totten SM, Feasley CL, Bermudez A, Pitteri SJ. Parallel Comparison of N-Linked Glycopeptide Enrichment Techniques Reveals Extensive Glycoproteomic Analysis of Plasma Enabled by SAX-ERLIC. J Proteome Res. 2017 Mar 3;16(3):1249-1260. Epub 2017 Feb 15


Acquisitions

SampleSpectrum FileLC Cal. SlopeLC Cal. Int.Trans. Groups
HEK293 CellsMS_47_UO1_HEK_1_1n2.506-32.7084
HEK293 CellsMS_47_UO1_HEK_1_2n2.375-30.1519
HEK293 CellsMS_47_UO1_HEK_2_1n2.399-30.31417
HEK293 CellsMS_47_UO1_HEK_2_2n2.346-29.6491
Human SerumDataMS_UO1_SWATH_11202019_Erlich105.086-89.1461
Human SerumDataMS_UO1_SWATH_11202019_Erlich35.36-87.78413
Human SerumDataMS_UO1_SWATH_11202019_Erlich45.757-103.91331
Human SerumDataMS_UO1_SWATH_11202019_Erlich55.086-89.146106
Human SerumDataMS_UO1_SWATH_11202019_Erlich65.048-89.45566
Human SerumDataMS_UO1_SWATH_11202019_Erlich75.463-109.178
Human SerumDataMS_UO1_SWATH_11202019_Erlich85.153-103.24765
Human SerumDataMS_UO1_SWATH_11202019_Erlich_W45.975-112.532
Human SerumDataMS_UO1_SWATH_11202019_Erlich_W55.198-91.95118
Human SerumDataMS_UO1_SWATH_11202019_Erlich_W65.823-117.8499
Human SerumDataMS_UO1_SWATH_11202019_Erlich_W84.058-63.6151
Human SerumMS_35_Frac_plasma_run01_fr10_012.553-31.911109
Human SerumMS_35_Frac_plasma_run01_fr11_012.53-30.4461
Human SerumMS_35_Frac_plasma_run01_fr12_012.534-30.75829
Human SerumMS_35_Frac_plasma_run01_fr13_012.782-61.0878
Human SerumMS_35_Frac_plasma_run01_fr14_012.489-30.3644
Human SerumMS_35_Frac_plasma_run01_fr1_012.483-29.9781
Human SerumMS_35_Frac_plasma_run01_fr2_012.47-29.9691
Human SerumMS_35_Frac_plasma_run01_fr3_012.478-30.0082
Human SerumMS_35_Frac_plasma_run01_fr4_012.468-29.93
Human SerumMS_35_Frac_plasma_run01_fr5_012.489-29.87627
Human SerumMS_35_Frac_plasma_run01_fr6_012.516-30.73583
Human SerumMS_35_Frac_plasma_run01_fr7_012.472-30.30673
Human SerumMS_35_Frac_plasma_run01_fr8_012.608-36.1291
Human SerumMS_35_Frac_plasma_run01_fr9_012.535-30.528118
Human SerumMS_35_Frac_plasma_run02_fr12.473-29.8961
Human SerumMS_35_Frac_plasma_run02_fr102.533-30.934106
Human SerumMS_35_Frac_plasma_run02_fr112.539-31.212124
Human SerumMS_35_Frac_plasma_run02_fr122.527-31.22108
Human SerumMS_35_Frac_plasma_run02_fr132.515-30.45883
Human SerumMS_35_Frac_plasma_run02_fr142.543-32.36925
Human SerumMS_35_Frac_plasma_run02_fr152.491-29.85913
Human SerumMS_35_Frac_plasma_run02_fr162.499-29.917
Human SerumMS_35_Frac_plasma_run02_fr172.598-34.135244
Human SerumMS_35_Frac_plasma_run02_fr22.485-30.4172
Human SerumMS_35_Frac_plasma_run02_fr42.52-31.1671
Human SerumMS_35_Frac_plasma_run02_fr52.578-32.14739
Human SerumMS_35_Frac_plasma_run02_fr62.523-30.32942
Human SerumMS_35_Frac_plasma_run02_fr72.473-29.51990
Human SerumMS_35_Frac_plasma_run02_fr82.51-30.60384
Human SerumMS_35_Frac_plasma_run02_fr92.554-32.16896
Human SerumMS_48_UO1_Erlic_E102.587-39.73136
Human SerumMS_48_UO1_Erlic_E112.475-36.047
Human SerumMS_48_UO1_Erlic_E122.46-35.9072
Human SerumMS_48_UO1_Erlic_E132.654-43.33617
Human SerumMS_48_UO1_Erlic_E142.521-38.4336
... further results