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Updated: Jan 4, 2026
Colorectal Cancer Cell Surface Protein Profiling Using an Antibody Microarray and Fluorescence Multiplexing
Published on: September 25, 2011
Sensitive glycoprofiling of insulin-like growth factor receptors isolated from colon tissue of patients with
Dragana Robajac1, Martina Križáková2, Romana Masnikosa1
1Institute for the Application of Nuclear Energy (INEP), University of Belgrade, Belgrade, Serbia.
Abstract:
Glycosylation of cell receptors influences their function and development of tumour induces changes in glycosylation. Cell growth depends on the activation of receptors which bind growth factors and the insulin-like growth factor (IGF) receptors are among the most important ones. Usually, only small quantities of isolated receptors are available thus there is a need of suitable assay to study receptors glycosylation. Therefore, we developed a lectin-based reverse-phase protein microarray method for screening the glycosylation pattern of receptors in picomolar (pM) concentrations. The method was applied to glycoprofile IGF1 and IGF2 receptors and the solubilised membrane proteins isolated from tumour and non-tumour colon tissue of patients with colorectal cancer. We found that common to both receptors was partial overlapping of the major glycan structures with those present in the entire glycome of membrane proteins. In contrast, receptors possess higher level of α2,3 sialic acid residues and lower level of tri-/tetra-antennary complex type N-glycans and terminal mannose in high-mannose structures. Increased levels of fucosylation and branched mannose structures were observed in both receptors derived from tumour tissue compared to non-tumour tissue. The described method enabling glycan analysis of receptors has a big application potential in e.g. biomarker research, biology and diagnostics.
Insights
This study introduces a new microarray method to analyze cell receptor glycosylation using small samples. The method revealed distinct glycan patterns in insulin-like growth factor receptors from tumor tissues, offering potential for biomarker research.
Area of Science:
- Glycobiology
- Cancer Biology
- Biomarker Discovery
Background:
- Cell receptor glycosylation is crucial for function and is altered in tumors.
- Insulin-like growth factor (IGF) receptors are key for cell growth and are often dysregulated in cancer.
- Limited availability of receptors necessitates sensitive assays for studying glycosylation.
Purpose of the Study:
- To develop a sensitive lectin-based protein microarray for analyzing receptor glycosylation at picomolar concentrations.
- To investigate and compare the glycosylation patterns of IGF1 and IGF2 receptors in colorectal cancer tissues.
- To identify potential glycan-based biomarkers for colorectal cancer.
Main Methods:
- Development of a lectin-based reverse-phase protein microarray assay.
- Application of the assay to glycoprofile IGF1 and IGF2 receptors and membrane proteins from tumor and non-tumor colon tissues.
- Quantitative analysis of glycan structures, including sialic acid, N-glycans, and mannose residues.
Main Results:
- The developed method can screen receptor glycosylation at picomolar concentrations.
- IGF1 and IGF2 receptors share some glycan structures with the overall membrane protein glycome.
- Receptors from tumor tissues show increased fucosylation and branched mannose structures compared to non-tumor tissues.
- Specific differences in sialic acid and N-glycan structures were observed between receptors and the general membrane glycome.
Conclusions:
- The lectin-based microarray is a powerful tool for analyzing receptor glycosylation with limited sample amounts.
- Alterations in IGF receptor glycosylation are associated with colorectal cancer development.
- The identified glycan changes represent potential biomarkers for cancer diagnostics and research.
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