Multiple Reaction Monitoring for the Quantitation of Serum Protein Glycosylation Profiles: Application to Ovarian Cancer

Abstract

Ascites fluid is present in over 30% of diagnosed ovarian cancer (OC) cases and is produced when tumor cells metastasize into the peritoneal cavity. Because of increased interest in the immune status of ovarian cancer patients who present with metastatic ascites and the potential role that glycosylation plays with functions of the different immunoglobulins such as complement-dependent and antibody-dependent cytotoxicity largely through the activity of Fc receptors. Immunoglobulins IgA and IgG were affinity isolated from OC ascites fluids, N-glycans were released and analyzed while site specific glycan analyses of glycopeptides was performed using tandem MS/MS with nano-LC Chip/QTOF MS. Results revealed the heterogeneity of N-glycans present in each glycosylation site of IgA1, IgA2 and IgG. Interestingly, fucosylated glycans were present in certain sites while other sites contained no fucosylated glycans. Highly sialylated N-glycans were present in other IgA sites suggesting different functional roles for each specific site.

Dramatically changed immune‐related molecules as early diagnostic biomarkers of non‐small cell lung cancer

“…ORM1 is an acute phase protein in humans and mice with inflammatory and immunomodulatory properties, the serum concentration of which rises two to five times during an acute phase response . Increasing expression of ORM1 has been reported to be used as a biomarker in various advanced cancers, such as bladder cancer , ovarian cancer and lung cancer , in studies that have focused on analysing late‐stage serum and urine samples with mass spectrometry. Zhang et al .…”

“…ORM1 is an acute phase protein in humans and mice with inflammatory and immunomodulatory properties, the serum concentration of which rises two to five times during an acute phase response [34,35]. Increasing expression of ORM1 has been reported to be used as a biomarker in various advanced cancers, such as bladder cancer [23], ovarian cancer [36] and lung cancer [37], in studies that have focused on analysing late-stage serum and urine samples with mass spectrometry. Zhang et al [38] previously identified that increased ORM1 acts as an early-detection protein biomarker of lung cancer in urine samples in KRas-driven lung cancer mouse models, which was validated in 16 human cancer urine samples and in tumour-bearing mice.…”

N-Glycosylation profiling of intact target proteins by high-resolution mass spectrometry (MS) and glycan analysis using ion mobility-MS/MS

“…Quantification of TFN glycoforms is commonly carried out at the glycopeptide level by triple quadrupole MRM experiments. 48,49 Compared to these methods, our method showed similar performances in terms of detection limits (ranging from 0.5 to 0.9 µg mL −1 ), precision, accuracy, and linearity. An example of quantification of TFN at the intact protein level was performed by LC-ICP-MS, focusing on the different sialoforms present on the protein.…”

Global and site‐specific analysis of protein glycosylation in complex biological systems with Mass Spectrometry

“…Due to the importance of protein glycosylation in biological systems, glycoproteomics has been extensively applied to disease-related research (Whelan et al, 2009;Hwang et al, 2010;Song et al, 2014;Reiding et al, 2017;Yang et al, 2017b;Kailemia et al, 2018), such as hepatitis, cancer (Dai et al, 2011;Liu et al, 2014;Kailemia et al, 2017;Miyamoto et al, 2018;Veillon et al, 2018), and infectious diseases (Vigerust, 2011;Go et al, 2017). Glycosylation can change chemical and physical properties of proteins, and regulate their binding and interactions with ligands or extracellular matrix.…”