Proteins & Peptides Product-related Impurity Analysis

In addition to process impurities, the isolation and characterization of product-related impurities is also an important aspect of process validation and batch-release testing for protein and peptide biopharmaceuticals. In accordance with the ICH Q6B guideline, CD Formulation's impurity analysis team applies a range of highly sensitive analytical techniques to characterize and test any possible product-related impurities, i.e., molecular variants of the desired product, including truncations, aggregates, modified forms, and certain degradation products.

What is Product-related Impurity?

The ICH Q6B guideline states that biopharmaceutical product-related impurities are mainly various molecular variants in the drug that are inconsistent with the function of the target product, among which the most common variants are products of chemical modification type variations and degradation products and aggregates of the target product.

Product-related impurities should be carried out in the early stages of product development. Complementary chromatography and electrophoresis methods with UV detection have been used to monitor therapeutic protein variants, but these methods are about to be replaced by multi-attribute methods (MAMs) that mainly use mass spectrometry as a detection tool. Using a single LC-MS or LC-MS/MS method can not only monitor but also characterize several of these variants or impurities, which will bring stronger identification capabilities.

Explore Our Proteins & Peptides Product-related Impurity Analysis

At CD Formulation, our scientists are experts in identifying product-related impurities for protein/peptide therapeutics. Leveraging expertise from decades of analytical experience, we are able to develop and customize solutions to identify and detect any possible product-related impurities.

We offer a range of analytical solutions to monitor impurities in your biopharmaceutical products in compliance with ICH Q6B and EMA guidelines. All of our methods can be validated to your specific project requirements and applied to process validation or testing of drug products or drug substance batches. These services are GMP-compliant and can be validated in accordance with ICH guidelines.

Our experienced scientists perform detailed characterization using a variety of techniques including MALDI-MS, LC-MSMS, HPLC, IR, NMR, and fluorescence methods.

We provide the following product-related impurity analysis services, including but not limited to:

Fig.1 Protein/peptide product-related impurity analysis. (CD Formulation)

Truncated Forms (Truncation, Fragmentation) Analysis

Protein biopharmaceuticals often break during production, preparation, and storage, resulting in shortened proteins. The truncation of some proteins may not only change their biological activity and in vivo pharmacokinetic characteristics but also become stronger antigens in the body, causing immune responses and affecting the safety and efficacy of the drug.

Our scientists use enzymes or chemicals to cleave peptides, and then characterize them by HPLC or SDS-PAGE. Our mass spectrometry experts also perform peptide mapping to obtain useful information about variants.

Incorrect Modification Forms (Disulfide, Oxidation/Deamidation, Glycosylation) Analysis

Precursors are inactive and often require a series of post-translational processing to become functional mature proteins. There are many types of processing, such as removal of N-terminal fMet or Met, disulfide bond formation, chemical modification, shearing, phosphorylation, methylation, acetylation, glycosylation, etc.

Our scientists use various chromatography, electrophoresis, or other related analytical methods, such as HPLC, MS, circular dichroism (CD), isoelectric focusing (IEF), peptide mapping, and nuclear magnetic resonance(NMR), to determine various incorrect modification forms, such as deamidation, isomerization, mismatched S-S connection, glycosylation, and phosphorylation.

Aggregation (Single/Multi-aggregate, Sub-visible Particles) Analysis

During the production, preparation, and storage of protein biopharmaceuticals, some forced denaturation conditions may be encountered, such as excessive acidity, heat denaturation, etc., which can easily cause protein entanglement and form aggregates.

Our scientists have extensive experience in determining the presence of aggregates, such as dimers and higher multiples of products. We can separate these aggregates from the desired product and perform quantitative analysis by size exclusion high-performance liquid chromatography (SEC-HPLC) or capillary electrophoresis.

Degradation Products Analysis

Our impurity experts are able to characterize a number of degradation products generated during manufacturing and/or storage and determine their levels according to appropriately established acceptance criteria.

Our Expertise

To meet regulatory agency requirements for product impurities, our portfolio of product-related impurity testing services is structured based on size variants, charge variants, or bioactivity.

• Physicochemical property testing: Many different physicochemical properties of proteins/peptides such as pH, concentration, and osmolality need to be tested during the manufacturing process. These properties are all necessary characteristics for manufacturing a consistent drug product.
• Charge variants: Charge variants are the result of post-translational modifications during manufacturing as well as chemical modifications during purification and storage. This impurity is a key factor affecting the bioactivity and pharmacokinetics of protein/peptide therapeutics. Our scientists use cation exchange chromatography (CEX) and capillary isoelectric focusing (cIEF) to characterize this class of impurities.
• Size distribution: Size variants, such as aggregates and fragments, can affect immunogenicity and potency, so it is important to monitor their presence. We use size exclusion chromatography (SEC), dynamic light scattering (DLS), and capillary electrophoresis sodium dodecyl sulfate gel (CE-SDS) to assess size distribution.
• Molecular weight analysis: Molecular weight is the primary indicator of protein/peptide therapeutic identity. It is often assessed through intact mass (IM) analysis.
• Post-translational modification (PTM) analysis: Understanding and controlling any modifications that occur during manufacturing helps produce consistent protein/peptide therapeutics. We use peptide mapping, LC-MS, or LC-MS/MS techniques to identify and quantify modifications, ensuring that the quality of the final product meets the requirements. By monitoring modification levels, we are able to optimize the manufacturing process and minimize any heterogeneity that may arise.

Available Product-related Impurity Testing Items

Why Choose Us for Proteins & Peptides Product-related Impurity Analysis?

• Our team of scientists and researchers have many years of experience in analyzing proteins and peptides and their associated impurities and possess the knowledge and skills required to accurately identify and quantify impurities in these complex molecules.
• Our laboratory is equipped with the latest technologies and instrumentation to perform comprehensive testing for any product-related impurity.
• We understand that each protein and peptide molecule is unique, and we tailor our analytical techniques to the specific characteristics of each molecule. This ensures that we provide the most comprehensive and relevant impurity analysis for your specific product.
• We are well-versed in the regulatory requirements for protein and peptide analysis, including those set forth by the FDA and other regulatory agencies. Our analytical reports are thorough and meet all necessary guidelines.
• We offer flexible testing options to meet the needs of your different projects.

Publication

Product-related impurity analysis is a critical component in ensuring the safety, efficacy, and consistency of therapeutic proteins and peptides. Please feel free to contact us if you are interested in our services. Learn how CD Formulation can help you conduct product-related impurity testing and verify the safety of your products in relation to regulatory requirements.

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.