For the quantification of the two distinct forms of AGT in the

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The second method was hypothesis generating and aimed to detect glycoprotein fold changes between different disease conditions using a simple and cost effective conventional LC MS/MS workflow.For both methods, a reproducible workflow for efficient glycoprotein/AGT extraction from human plasma was developed by coupling ConA lectin affinity chromatography with reversed phase solid phase extraction fractionation (RP SPE). Analysis of the enzymatically digested proteins was conducted using targeted LC MS/MS working under the multiple reaction monitoring mode. For the quantification of the two distinct forms of AGT in the plasma (the sulphydryl bridged oxidised form and the free thiol reduced form), a differential alkylation strategy was coupled with targeted LC MS/MS to recognise and quantify the cysteine (Cys) peptides involved in the redox switch of AGT.

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