Sentences with phrase «immunoaffinity depletion»

For label - free quantification analyzing unfractionated CSF samples (individual patient samples with insufficient volume (protein content) for immunoaffinity depletion and SCX fractionation), the LTQ - Orbitrap Velos mass spectrometer was operated in the data - dependent mode with full scan MS spectra (m / z 400 — 2000) acquired in the LTQ - Orbitrap Velos with resolution of 60,000 at m / z 400 (accumulation target: 1,000,000).

Limited volumes of the individual samples reduced the sample preparation options (i.e., immunoaffinity depletion and SCX fractionation), and hence resulted in less depth of proteome coverage than possible with the pooled samples, where approximately 20 ml were available for depletion and fractionation.

Pooled cerebrospinal fluid (CSF) samples from nPTLS patients, CFS patients, and healthy volunteers were comprehensively analyzed using high - resolution mass spectrometry (MS), coupled with immunoaffinity depletion methods to reduce protein - masking by abundant proteins.

CSF proteins (from the immunoaffinity depletion processed pooled samples and the individual samples without immunoaffinity depletion processing) were digested with trypsin and cleaned up with SPE C18 columns as previously described [10].

Coupling immunoaffinity depletion with strong cation exchange (SCX) fractionation further reduces sample complexity, and allowed for the in - depth analysis of the CSF proteomes.

Pooling samples provided sufficient protein mass for effective downstream proteomics analysis following immunoaffinity depletion of the 14 most abundant proteins present (representing approximately 95 % of the total protein mass in CSF), reducing the dynamic range of protein concentrations present in CSF, where proteins with highest concentrations mask proteins at lower concentrations from detection.

Equal aliquots from individual CSF samples were pooled to provide sufficient volume for extensive fractionation and two - dimensional LC coupled to tandem MS (2D - LC - MS / MS) analysis with immunoaffinity depletion from 30 women and 13 men (n = 43) who fulfilled the 1994 case definition for CFS [1].

Individual CSF samples from 14 of the 43 CFS subjects (aged 33 — 48 years with a median age of 43 years, 7 female and 7 male) were also used in direct LC - MS analysis (i.e., no MS / MS was performed) without immunoaffinity depletion.

Equal aliquots from individual CSF samples were pooled to provide sufficient volume for extensive fractionation and 2D - LC - MS / MS analysis with immunoaffinity depletion from 15 females and 10 males (n = 25) with nPTLS.

Individual CSF samples from another set of 10 healthy volunteers, age 37 — 44 years (median = 40) and 5 women and 5 men, were analyzed by LC - MS analysis without immunoaffinity depletion.