Quick Guide to Released Glycan Analysis

Released Glycan (N-linked), Released Glycan (O-linked), and Released Glycan (IgG) Workflows

These Released Glycan workflows all have four separate tabs: Samples, Glycan table, Glycan options, and Processing nodes. Please note the default setting for these analyses are set to process in negative mode.

Create a Workflow

Click to launch any of the above mentioned workflows. In the figure below, the Released Glycan (N-linked, neg mode) workflow is highlighted.

Figure 1: Byos System Workflows with Released Glycan (N-linked, neg mode) highlighted

Drag and drop the sample file(s) to be processed into the Samples tab, as shown in the figure below. These can be from any vendor.

Figure 2: Drag and drop sample data files to be processed into the Samples tab

The user can load a Traces file, an MS/MS search result *.byrslt file, and has an option to set the digestion parameters. To load the trace file and MS/MS search result file, click within the box to activate the light blue “…” box, and upload the respective files. For setting the digestion parameter, use the dropdown menu to select the enzyme.

Figure 3: Click to activate the traces editor in TracesCsv or TracesMs

The user can associate traces with samples. To associate traces, click within the box to activate the light blue “…” box, as shown in the figure below.

Figure 4: Associate traces with samples. Traces are assigned to all samples by default

Each default workflow includes a table of common glycans. The user can choose to use the default list, add to it, or replace it entirely with a customized list with the Add from CSV file button.

Figure 5: Glycan table tab

If using a fluorescent tag, the user can set this in the Glycan options tab. Several commonly used tags are included by default; if an alternative mass is preferred, please use the Custom radio button. The user can also add adduct masses to be considered in calculations using the Add Adduct button.

Figure 6: Glycan options tab. Set the label, include adducts using the Add Adduct button

Click Create Project to start analysis and report generation. Please note Byos will save this type of project as a *.bmap file.

Figure 7: Click Create Project.

The user will be prompted to name and save the analysis file.

Figure 8: Name the project and save in a convenient location

Processing will begin. Once complete, the project and associated report will appear as additional tabs after the Workflows tab, as shown in the figure below.

Figure 9: Workflow tab, completed project, and associated report are available as separate tabs