engleski

Analysis of Fc-linked N-glycosylation of mouse IgG with nanoUPLC-ESI-MS

It is known that glycosylation of immunoglobulin G is important for its biological functions. Mouse models are often used in immunology, however detailed studies of mouse IgG glycome are scarce. In this study we test the reproducibility of the sample preparation procedure used for analysis of Fc- linked N-glycans on IgG isolated from mouse serum. To test technical variation introduced by each step starting from isolation of IgG we used replicates of BALB/c mouse serum sample. At each step 19 replicates were pooled together to remove technical variation introduced by previous steps, aliquoted and treated as individual samples for the rest of the procedure. Tryptic glycopeptides of Fc fragment of IgG in each sample were analyzed on nanoUPLC-ESI-MS. Using LaCyTools software we have identified analytes of interest and extracted their corresponding areas to test the reproducibility. We identified the step of IgG affinity isolation as introducing most of technical variation in the replication experiment and reproducibility of the whole sample preparation procedure to be acceptable for reliable quantification of glycopeptides.

IgG ; Fc-linked N-glycosylation ; nanoUPLC-ESI-MS ; mouse

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