LC-MS Based Host Residual Protein Analysis Service

Host cell proteins (HCPs), derived from the production cell lines used in biopharmaceuticals, include structural and secreted proteins of host cells. These proteins vary widely in properties such as isoelectric points, hydrophobicity, and molecular weight. As unintended components in bioproducts, HCPs pose risks of immunogenicity, impacting safety and drug stability. Regulatory bodies, such as the International Council for Harmonisation (ICH), require rigorous monitoring and control of residual HCPs. Traditional ELISA methods, often limited by the availability of specific antibodies, fail to detect all HCP varieties comprehensively.

 

MtoZ Biolabs utilizes Thermo Fisher’s Orbitrap Fusion Lumos mass spectrometer combined with nano-liquid chromatography, offering an LC-MS based service for detailed qualitative and quantitative analysis of HCPs, ensuring comprehensive coverage and precision in biopharmaceutical applications.

 

Analysis Workflow

1. Sample Preparation

Extract total proteins from host cells.

 

2. Protein Digestion

Treat samples with trypsin to generate peptide fragments.

 

3. LC-MS Analysis

Separated by LC, peptides are then analyzed using MS for detailed detection.

 

4. Data Analysis

Use advanced bioinformatics tools to analyze data, ensuring accuracy and reliability.

 

Service Advantages

Antibody Independence: Independent of specific antibodies, avoiding limitations associated with antibody availability, enabling the discovery of a broader range of HCPs.

 

High Sensitivity: Capable of detecting HCPs at extremely low concentrations, far surpassing the sensitivity of ELISA.

 

Extensive Dynamic Range: Suitable for detecting proteins at concentrations from very low to very high, with a broad dynamic range.

 

Fast and Accurate: Provides more rapid and accurate analysis results compared to ELISA.

 

Sample Results

Enhancing HCP Detection in Protein Therapeutics Using HILIC Enrichment and LC-MS/MS



Wang, Q. Y. et al. Anal. Chem. 2020.

 

SP3-Based HCP Monitoring in AAV-Based Gene Therapy Products Using LC-MS/MS

 



Smith, J. et al. Eur J Pharm Biopharm. 2023.

 

Deliverables

1. Experimental Procedures

2. Relevant Mass Spectrometry Parameters

3. Detailed Information on HCPs

4. Mass Spectrometry Images

5. Raw Data