N-Glycan Modification and Modification Site Analysis Service

N-glycoproteins are proteins modified by the addition of glycan chains through N-glycosylation on specific amino acid residues. This modification predominantly occurs in sequences containing asparagine (Asn) residues within the Asn-X-Ser/Thr motif, where X represents any amino acid except proline. N-glycans critically influence protein folding, stability, biological activity, and intercellular communication. For instance, glycan chains can alter protein's three-dimensional structure, affecting their molecular interactions. Furthermore, these chains protect proteins from enzymatic degradation and extend their circulatory half-life, which is crucial for pharmaceutical applications. N-glycan modifications involve the attachment of glycan chains at specific protein sites, typically at asparagine residues within the Asn-X-Ser/Thr motif. Analyzing these modifications and their specific sites is vital for understanding protein functions and interactions within biological systems. In biopharmaceuticals, such analysis is essential for optimizing drug efficacy and safety, ensuring proper pharmacokinetics and pharmacodynamics. It also plays a crucial role in verifying the glycosylation profiles of biosimilars relative to original therapeutics, critical for their approval and clinical efficacy.

 

O-glycosylation, a prevalent post-translational modification, significantly influences protein structure and functionality. According to the ICH Q6B guidelines, it is mandatory to determine the glycosylation levels, sites, and types. Leveraging sophisticated mass spectrometry platforms and extensive expertise, MtoZ Biolabs delivers exceptional services in N-glycan modification and site analysis.

 



Cao, L. et al. Nat Protoc. 2018.

Figure 1. The Workflow of Site-specific Analysis of Glycoprotein N-glycan Processing

 

Analysis Workflow

1. Extracting and Purifying the Target Protein

2. Enzymatic Digestion to Release the N-glycans

3. Glycan Separation

4. Structural Analysis of Glycans

5. Site-specific Glycan Analysis

6. Data Analysis and Interpretation

 

Service Advantages

1. Tailored and Specialized Service: MtoZ Biolabs provides customized solutions tailored to the specific needs of various biopharmaceuticals and clients, including comprehensive glycan profiling and both quantitative and qualitative structural analyses.

2. Advanced Technological Integration: Our cutting-edge technologies such as mass spectrometry, liquid chromatography, and capillary electrophoresis ensure high precision and throughput in N-glycan analysis, guaranteeing reliable and consistent results.

 

Applications

1. Enhancing Drug Safety and Efficacy

N-glycan modifications are crucial quality attributes in biopharmaceuticals that influence the drugs' biological activity, stability, and safety. Precise analysis of glycosylation sites and glycan structures ensures the consistency and predictability of biopharmaceutical products, enhancing their therapeutic efficacy and safety.

 

2. Production Optimization

Continuous monitoring and analysis of N-glycan variations during manufacturing processes enable process optimization, enhancing efficiency and quality, which are essential for cost reduction and production cycle minimization.

 

3. Exploring Drug Mechanisms

Analysis of N-glycan modification sites enriches the understanding of drug mechanisms, particularly for biopharmaceuticals that function through interactions with cell surface receptors. This insight is invaluable for drug refinement and development of new therapeutics.

 

Sample Requirements

Protein: Minimum of 100 μg.

Cells: At least 1x10⁷ cells.

Animal Tissue: Approximately 1 g.

Blood: 1 mL, which must be anticoagulated with EDTA.

Serum: Between 0.2 to 0.5 mL.

Urine: At least 2 mL.

Microbial Samples: 200 mg in dry weight.

 

Please note: Should you have any inquiries regarding sample submission, do not hesitate to reach out. Our expert team is available to assist you at all times.

 

Deliverables

1. Experimental Procedures

2. Relevant Mass Spectrometry Parameters

3. Detailed Information on N-Glycan Modification and Modification Site Analysis

4. Mass Spectrometry Images

5. Raw Data