Proteins & Peptides Particle and Aggregation Characterization

Protein aggregates and particles can form at any stage of the therapeutic protein/peptide development and manufacturing process, including bioprocessing, purification, formulation and packaging, and storage. As one of the key steps in the successful development and manufacturing of biopharmaceuticals, protein aggregation and particle characterization helps assess the extent of protein degradation, stability, and aggregate formation in solution. CD Formulation offers a wide range of analytical techniques for characterizing protein/peptide aggregation to gain an in-depth understanding of the aggregation of your product during development and manufacturing, thereby helping to optimize the manufacturing process and improve product quality and stability.

What is Protein Aggregation?

Protein aggregation is a biological phenomenon in which proteins aggregate and condense in solution to form large particles or aggregates. This phenomenon may cause the protein to lose biological activity, cause structural changes in the protein, or even impair its function. Protein aggregates or particles can be considered impurities or molecular variants, which may be caused by adverse conditions or specific environmental factors, such as changes in pH, temperature, ionic strength, reactions with excipients in the formulation, buffer concentration, or interactions with the container/closure system (such as elastic seals or glass delamination). Fig.1 shows the possible competitive aggregation mechanisms of proteins at low and high concentrations.

Fig.1 Possible competitive aggregation mechanisms of proteins at low and high concentrations. (Housmans JAJ, et al., 2021)

The potential aggregation mechanisms in the figure above determine the possible combinations of aggregate population size and concentration. Typically, aggregates include reversibly non-covalently and irreversibly covalently bound species, dimers, oligomers and higher multiples of the desired protein product, and can exist as small soluble particles ranging in size from a few nanometers to large sub-visible/visible particles up to micrometers.

Significance of Aggregation Characterization of Therapeutic Proteins And Peptides

Protein aggregation is one of the key challenges in the development of protein biotherapeutics. Aggregation of protein or peptide therapeutics can have serious impacts on product stability, biological activity, efficacy, quality, and patient safety. An obvious danger is that protein or peptide aggregation can increase the potential for immunogenicity, leading to adverse immune responses in patients. Therefore, it is very important to mitigate health risks during drug development by fully understanding the aggregation propensity of biomolecules and characterizing the aggregation state.

Explore Our Proteins & Peptides Particle and Aggregation Characterization Services

FDA regulations strongly recommend in-depth characterization of the number and type of aggregates and particles that may be present in biotherapeutics. At CD Formulation, our team of protein experts applies a range of protein aggregation and particle analysis techniques to detect and quantify any aggregates that may be present in therapeutic protein or peptide solutions, further supporting formulation development, quality control, stability studies, comparability, release testing, and aggregation studies.

Our protein or peptide aggregation characterization services are performed in accordance with the ICH Q6B guideline. Throughout the aggregation and particle analysis process, our team of experts combines multiple technical strategies such as differential scanning calorimetry (DSC), analytical ultracentrifugation (AUC) and light scattering techniques (DLS, SEC-MALS) to study the non-covalent structural aspects of proteins and the formation of aggregates, oligomers or particles in solution.

Fig. 2 Protein/peptide particle and aggregation characterization.(CD Formulation)

• For sub-visible particles, we use light obscuration (LO) technology, which is the primary compendial technology authorized by the USP. This technique is very effective for counting and sizing opaque particles.  Furthermore, flow imaging microscopy (FIM) is another compendial technique we use to analyze particle counting and sizing to complement LO. This technique is also a USP-recognized orthogonal technique (via USP <1788>).
• For submicron aggregates, we typically use size exclusion chromatography (SEC) and hydrophobic interaction chromatography (HIC) for aggregation analysis. The former separates particles based on hydrodynamics and can detect and quantify irreversible aggregates from oligomers to very high-order aggregates, often in combination with multi-angle static light scattering (SEC-MALS), while the latter separates proteins based on hydrophobicity in their native state and can often detect changes in protein structure and aggregates.
• Dynamic light scattering (DLS) can provide useful size and polydispersity information for monomers/oligomers and particles between 1 nm and 1 μm, thus measuring the particle size distribution of proteins in solution, particle size analysis, and zeta potential measurement. The size distribution obtained by DLS is biased towards larger particles.
• Analytical ultracentrifugation (AUC) can assess the homogeneity of protein/peptide solutions and qualitatively assess molecular weight and the presence of aggregates, ranging from a few kDa to MDalton. therefore, AUC can uniquely analyze multiple species in the same sample, while most other methods for aggregation analysis are limited due to the size range or resolution between different oligomers.

Our Technology Platforms

Why Choose Us for Proteins & Peptides Particle and Aggregation Characterization?

• Extensive experience in performing protein/peptide aggregation and particle analysis.
• Has multiple advanced technology platforms.
• Provides flexible experimental design and testing options can meet any customer's specific needs.
• Has Hundreds of successful project experience using protein/peptide aggregation and particle analysis to support biopharmaceutical product development.

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CD Formulation is designed to provide a powerful analytical tool to support formulation development, quality control, stability studies, comparability studies, etc., for protein and peptide biopharmaceutical products. Please feel free to contact us if you are interested in our services. We will provide you with the most professional advice and support to ensure the smooth launch and implementation of your project.

How It Works

STEP 1

Submit a service request describing your specific research or development need.

STEP 2

We'll email you to provide your quote and confirm order details if applicable.

STEP 3

Execute the project with real-time communication, and deliver the final report promptly.