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. 2018 Aug 10;9(9):889–894. doi: 10.1021/acsmedchemlett.8b00127

Figure 5.

Figure 5

Glycoform-focused quantitative analysis of therapeutic antibody based on the SRM channels made by using synthetic human IgG1 Fc fragments having a bisected G2 (1) and a biantennary G2 (2). (A,B) Calibration curves for glycopeptides 1 and 2 were made by plotting the results of TICs (Figure 4). (C,D) TICs in the SRM assay using 1.5 μL of tryptic digests of Herceptin (158 pmole) uncovered the peaks of Q3 (m/z 366.2) at 2.46 min and Q3 (m/z 2447.8) at 3.30 min representing the occurrence of the targeted fragments corresponding to the Fc glycopeptides 1 (left) and 2 (right), respectively. Because of the detection limit, calibration curve for 1 did not give an ideal linearity below 200 fmol, indicating that accuracy on analyte quantities should be assessed in the assayed concentration range.