Rapid Glyco-Qualitative Assessment of Recombinant Proteins Using a Fully Automated System
June 28th, 2024
A system for the automated and rapid analysis of glycans on proteins was previously developed. This article presents the protocol for "glyco-qualitative analysis," optimized for a broad spectrum of users, such as those engaged in analyzing glycan structures in biopharmaceuticals and other glycoconjugate materials.
Our scope is protein glycosylation. Glycan structures are involved in changes in the stability and function of biopharmaceuticals, and are altered along with host cell species and culture conditions. In some production conditions, allergenic glycan would be contaminated to a certain degree.
Our goal is to make user-friendly tools for such glyco-qualification. Mostly, LC and mass spectrometry. But for simple testing, lectin-based technologies are applicable.
Researchers have available many types of lectins with different specificities, and each lectin binds to some specific glycans. So, we can estimate glycan structures using multiuse of lectin-by-lectin blotting, lectin ELISA, and lectin microarray. Glycosylation occurs in functional proteins in our body.
Glycan structure is altered along with diseases. Nowadays, we have used several disease-specific glycosylations as diagnostic biomarkers, such as CA 19-9 and AFP-L3. Notably, new biomarker candidates have been discovered using lectin-based technologies.
For their validation, we need a glyco-qualification system. Glyco-qualification is hard work. We need to analyze hundreds or thousands of numbers in a short time.
However, there is no system to meet the needs. Our analytical method is the only one with a full automatic measurement process. It drastically reduces manpower and enables us to conduct effective validation.
Our work aims to further improve our rapid instrument, especially for quantitative measurement. After this, we expect to realize a unique diagnostic tool with a glyco-biomarker panel measured by our multilectin system. Hence, we are focusing on the discovery of disease-specific glycobiomarkers.
Overview
This article presents a protocol for glyco-qualitative analysis, focusing on the automated and rapid analysis of glycans on proteins. It aims to provide user-friendly tools for researchers analyzing glycan structures in biopharmaceuticals and glycoconjugate materials.
Key Study Components
Area of Science
- Biopharmaceuticals
- Glycobiology
- Analytical Chemistry
Background
- Protein glycosylation affects stability and function of biopharmaceuticals.
- Glycan structures can be altered by host cell species and culture conditions.
- Allergenic glycans may contaminate products under certain conditions.
- Lectin-based technologies are available for glycan analysis.
Purpose of Study
- To develop a glyco-qualification system for rapid analysis.
- To facilitate the discovery of disease-specific glycobiomarkers.
- To improve quantitative measurement capabilities in glycan analysis.
Methods Used
- Lectin-by-lectin blotting
- Lectin ELISA
- Lectin microarray
- Automated measurement processes
Main Results
- Development of a fully automated glyco-qualification system.
- Reduction of manpower needed for glycan analysis.
- Identification of new biomarker candidates using lectin technologies.
- Establishment of a glyco-biomarker panel for diagnostics.
Conclusions
- The automated system significantly enhances the efficiency of glycan analysis.
- It supports the validation of disease-specific glycosylation biomarkers.
- Future work aims to refine quantitative measurement tools.
