Antibody Impurity Analysis

Monoclonal antibodies (mAbs) have been successfully employed to target a wide range of therapeutic areas over the last two decades. The production of mAbs is typically from expression systems such as bacteria, yeast, mammalian, insect, and plants, which generate a variety of process-related purities. In the meantime, product-related impurities can also be produced through the manufacturing and purification process. These impurities can potentially compromise both drug efficacy and safety (e.g. eliciting unwanted immunogenic responses), thereby making it necessary to give an extensive characterization of antibody impurities with highly sensitive and specific analytical techniques.

Process-Related Residual & Product-Related Impurity

The production of mAb therapeutics involves a variety of steps, including upstream and downstream processes. Process-related impurities (e.g., raw materials, host cell protein, host cell DNA, adventitious and endogenous viruses, media components, leachables, and chemical additives) and product-related impurities (e.g., aggregates, breakdown products, product variants) can be derived from any stage of the process. These impurities must be monitored and effectively removed because they may cause adverse reactions. To ensure that the level of these impurities is within safe limits, highly sensitive analytical tools are needed for their detection and quantification. Due to the variety of impurities, Creative Biolabs applies a range of highly specific and sensitive techniques to analyze the impurities of mAb therapeutic products.

Analysis of Process-Related Impurities

Process-related impurities include cell substrates (e.g., host cell proteins (HCPs), host cell DNA (HC-DNA)), cell culture (e.g., inducers, antibiotics, and media components), or chromatographic media used in purification, solvents and buffer components.

• ELISA kits or mass spectrometry (MS)-based techniques are utilized for HCP analysis while PCR is often used for HC-DNA analysis.
• HPLC or LC-MS/MS techniques are applied to determine antibiotics (e.g., kanamycin, tobramycin, gentamycin), antifoam agents, and surfactants.
• Capillary electrophoresis sodium dodecyl sulfate (CE-SDS) is widely used to determine mAb purity. It provides automated, quantitative purity data on both intact and reduced mAbs.
• Other impurities including inducers (e.g., IPTG), serum, and process enhancing agents can also be determined.

Analysis of Product-Related Impurities

Product-related impurities are molecular variants of the desired product, including truncated and other modified forms, aggregates, precursors and certain degradation products arising during manufacture and/or storage.

• For instance, we can analyze the disulfide bridge scrambling, conformational impurities, and C-terminal integrity by peptide mapping using LC-MS/MS.
• We can also combine chromatographic, electrophoretic, and other analytical methods with MS-based methods.
• Moreover, we analyze the aggregation of mAb with techniques such as analytical ultracentrifugation (AUC), SEC/SEC-MALLS, dynamic light scattering (DLS).
• Furthermore, we analyze post-translational modifications (PTMs) variants including N-terminal cyclization, oxidation, glycation, glycosylation, etc.

Features of Our Services

• Perform detailed characterization using a diverse range of analytical techniques
• Investigate the effect of impurities on antibody efficacy and performance using our established antibody function assays
• Eliminate the unwanted proteins if needed
• Tailored analytical services to meet different client requirements

As a world-leading CRO and recognized expert for antibody development, characterization, analysis, and manufacturing, Creative Biolabs are equipped with modern technologies and instruments for your therapeutic antibody development projects. We can give a comprehensive characterization of your candidate mAbs in terms of their structure, modifications, aggregation, stability, and purity. If you are interested in our service, please contact us to discuss your requirements.

For Research Use Only.