GlycoFibroTyper: A Novel Method for the Glycan Analysis of IgG and the Development of a Biomarker Signature of Liver Fibrosis.
GlycoFibroTyper: A Novel Method for the Glycan Analysis of IgG and the Development of a Biomarker Signature of Liver Fibrosis.
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DOI：
10.3389/fimmu.2022.797460
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发表时间：
2022
影响因子：
7.3
通讯作者：
Mehta AS
中科院分区：
文献类型：
--
作者：
Scott DA;Wang M;Grauzam S;Pippin S;Black A;Angel PM;Drake RR;Castellino S;Kono Y;Rockey DC;Mehta AS
我们的团队最近开发了 GlycoTyper 测定，这是一种简化的抗体捕获载玻片阵列方法，可直接分析捕获的血清糖蛋白（包括免疫球蛋白 G (IgG)）的 N-聚糖。该方法仅需要几微升血清，并采用简化的处理方案，在分析前无需纯化或糖修饰。在此方法中，抗体捕获的糖蛋白用肽 N-糖苷酶 F (PNGase F) 处理，释放 N-聚糖，用于 MALDI 成像质谱 (IMS) 检测。由于大型纤维化和肝硬化患者队列中的 IgG 发生了 N 连接聚糖的改变，我们利用这种新方法在 106 名活检证实肝纤维化患者的队列中检查了总 IgG 以及以前从未检查过的 IgG1、IgG2、IgG3 和 IgG4 的糖基化。患者被分为无纤维化证据（41 名无肝病或 0 期纤维化患者）、早期纤维化（10 名 METAVIR 1 期患者和 18 名 METAVIR 2 期患者）或晚期纤维化（6 名 METAVIR 3 期纤维化患者和 37 名 METAVIR 4 期纤维化患者（肝硬化））。观察到糖基化的几个主要改变，将患者分类为无纤维化（敏感性为 92%，特异性为 90%）、早期纤维化（敏感性为 84%，特异性为 90%）或显着纤维化（敏感性为 94%，特异性为 90%）。
Our group has recently developed the GlycoTyper assay which is a streamlined antibody capture slide array approach to directly profile N-glycans of captured serum glycoproteins including immunoglobulin G (IgG). This method needs only a few microliters of serum and utilizes a simplified processing protocol that requires no purification or sugar modifications prior to analysis. In this method, antibody captured glycoproteins are treated with peptide N-glycosidase F (PNGase F) to release N-glycans for detection by MALDI imaging mass spectrometry (IMS). As alterations in N-linked glycans have been reported for IgG from large patient cohorts with fibrosis and cirrhosis, we utilized this novel method to examine the glycosylation of total IgG, as well as IgG1, IgG2, IgG3 and IgG4, which have never been examined before, in a cohort of 106 patients with biopsy confirmed liver fibrosis. Patients were classified as either having no evidence of fibrosis (41 patients with no liver disease or stage 0 fibrosis), early stage fibrosis (10 METAVIR stage 1 and 18 METAVIR stage 2) or late stage fibrosis (6 patients with METAVIR stage 3 fibrosis and 37 patients with METAVIR stage 4 fibrosis (cirrhosis)). Several major alterations in glycosylation were observed that classify patients as having no fibrosis (sensitivity of 92% and a specificity of 90%), early fibrosis (sensitivity of 84% with 90% specificity) or significant fibrosis (sensitivity of 94% with 90% specificity).