Affinity chromatography assisted comprehensive phosphoproteomics analysis of human saliva for lung cancer

Wang, Chentong; Qian, Liqiang; Ji, Liyun; Li, Sha; Wahid, Amir; Jiang, Xiaoteng; Sohail, Amir; Ji, Yin; Zhang, Yan; Wang, Peng; Xiao, Hua

doi:10.1016/j.aca.2020.03.043

Cited by 21 publications

(19 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The chelation between the phosphate groups of phosphopeptides and the metal ions such as Fe (III), Al (III), Ga (III), and Ti(IV) immobilized on the sorbent surface is a typical interaction in the enrichment of phosphopeptides by IMAC [42][43][44]. The most commonly used ligands for the immobilization of metal ions are iminodiacetic acid (IDA) or nitrilotriacetic acid (NTA).…”

Section: Immobilized Metal Affinity Chromatographymentioning

confidence: 99%

“…In the last decade, there has been an increasing trend in the enrichment of biomarkers with IMAC in various biotechnological and biomedical applications before mass spectrometric analysis for their detection in biological samples [7, 9, 42–44, 47–50, 14,15,51–60]. IMAC has become an important technique in the purification of histidine‐tagged proteins and phosphoproteins in recent years.…”

Section: Bioaffinity Chromatography Techniquesmentioning

confidence: 99%

See 1 more Smart Citation

Recent trends in sorbents for bioaffinity chromatography

Kip

Hamaloğlu

Demir

et al. 2021

J of Separation Science

View full text Add to dashboard Cite

Isolation or enrichment of biological molecules from complex biological samples is mostly a prerequisite in proteomics, genomics, and glycomics. Different techniques have been used to advance the efficiency of the purification of biological molecules. Bioaffinity chromatography is one of the most powerful technique that plays an important role in the isolation of target biological molecules by the specific interactions with ligands that are immobilized on different support materials. This review examines the recent developments in bioaffinity chromatography particularly over the past 5 years in the literature. Also properties of supports, immobilization techniques, types of binding agents, and methods used in bioaffinity chromatography applications are summarized.

show abstract

Section: Immobilized Metal Affinity Chromatographymentioning

confidence: 99%

Section: Bioaffinity Chromatography Techniquesmentioning

confidence: 99%

Recent trends in sorbents for bioaffinity chromatography

Kip

Hamaloğlu

Demir

et al. 2021

J of Separation Science

View full text Add to dashboard Cite

show abstract

“…18 Furthermore, as shown in our previous study, through Ti 4+ -IMAC affinity chromatography enrichment and phosphoproteomics analysis, 796 unique phosphopeptides corresponding to 517 phosphoproteins were characterized in human saliva, and 175 phosphorylation sites were differently expressed in the lung cancer group. 19 Due to the high macroheterogeneity and microheterogeneity of glycans, intact glycopeptides analysis is becoming an urgent need to detect unique site-specific glycoforms associated with cancers. 20 Of note is that glycopeptides are usually in low abundance, and they should be enriched prior to glycoproteomics analysis.…”

Section: Introductionmentioning

confidence: 99%

Integrated N-glycoproteomics Analysis of Human Saliva for Lung Cancer

Wang

Jiang

et al. 2022

J. Proteome Res.

Self Cite

View full text Add to dashboard Cite

Aberrant protein N-glycosylation is a cancer hallmark, which has great potential for cancer detection. However, large-scale and in-depth analysis of N-glycosylation remains challenging because of its high heterogeneity, complexity, and low abundance. Human saliva is an attractive diagnostic body fluid, while few efforts explored its N-glycoproteome for lung cancer. Here, we utilized a zwitterionic–hydrophilic interaction chromatography-based strategy to specifically enrich salivary glycopeptides. Through quantitative proteomics analysis, 1492 and 1234 intact N-glycopeptides were confidently identified from pooled saliva samples of 10 subjects in the nonsmall-cell lung cancer group and 10 subjects in the normal control group. Accordingly, 575 and 404 N-glycosites were revealed for the lung cancer group and normal control group. In particular, 154 N-glycosites and 259 site-specific glycoforms were significantly dysregulated in the lung cancer group. Several N-glycosites located at the same glycoprotein and glycans attached to the same N-glycosites were observed with differential expressions, including haptoglobin, Mucin-5B, lactotransferrin, and α-1-acid glycoprotein 1. These N-glycoproteins were mainly related to inflammatory responses, infectious diseases, and cancers. Our study achieved comprehensive characterization of salivary N-glycoproteome, and dysregulated site-specific glycoforms hold promise for noninvasive detection of lung cancer.

show abstract

“…Abnormal N-glycosylation and phosphorylation of proteins are closely related to the occurrence and development of many diseases, especially cancer ( Zhu et al, 2019 ; Mahoney et al, 2021 ). In recent years, many studies have found N-glycoproteins and phosphoproteins as potential biomarkers or drug targets for cancer ( Wang et al, 2020 ; Lin et al, 2021 ). Our tandem enrichment strategy was applied in a lung cancer study and led to the discovery of 249 and 486 differentially expressed N-glycosylated and phosphorylated proteins between cancer and normal tissues, respectively.…”

Section: Introductionmentioning

confidence: 99%

A New Strategy for High-Efficient Tandem Enrichment and Simultaneous Profiling of N-Glycopeptides and Phosphopeptides in Lung Cancer Tissue

Yang

Liu

et al. 2022

Front. Mol. Biosci.

View full text Add to dashboard Cite

N-glycosylation and phosphorylation, two common posttranslational modifications, play important roles in various biological processes and are extensively studied for biomarker and drug target screening. Because of their low abundance, enrichment of N-glycopeptides and phosphopeptides prior to LC–MS/MS analysis is essential. However, simultaneous characterization of these two types of posttranslational modifications in complex biological samples is still challenging, especially for tiny amount of samples obtained in tissue biopsy. Here, we introduced a new strategy for the highly efficient tandem enrichment of N-glycopeptides and phosphopeptides using HILIC and TiO2 microparticles. The N-glycopeptides and phosphosites obtained by tandem enrichment were 21%–377% and 22%–263% higher than those obtained by enriching the two PTM peptides separately, respectively, using 160–20 μg tryptic digested peptides as the starting material. Under the optimized conditions, 2798 N-glycopeptides from 434 N-glycoproteins and 5130 phosphosites from 1986 phosphoproteins were confidently identified from three technical replicates of HeLa cells by mass spectrometry analysis. Application of this tandem enrichment strategy in a lung cancer study led to simultaneous characterization of the two PTM peptides and discovery of hundreds of differentially expressed N-glycosylated and phosphorylated proteins between cancer and normal tissues, demonstrating the high sensitivity of this strategy for investigation of dysregulated PTMs using very limited clinical samples.

show abstract

Affinity chromatography assisted comprehensive phosphoproteomics analysis of human saliva for lung cancer

Cited by 21 publications

References 47 publications

Recent trends in sorbents for bioaffinity chromatography

Recent trends in sorbents for bioaffinity chromatography

Integrated N-glycoproteomics Analysis of Human Saliva for Lung Cancer

A New Strategy for High-Efficient Tandem Enrichment and Simultaneous Profiling of N-Glycopeptides and Phosphopeptides in Lung Cancer Tissue

Contact Info

Product

Resources

About