Proteins & Peptides Forced Degradation Studies

Forced degradation studies of proteins/peptides are an integral part of biopharmaceutical development. Understanding potential degradation pathways through forced or stress studies is an important factor in determining the critical quality attributes (CQAs) of protein therapeutics. This ensures that optimal quality control strategies are implemented to monitor the continued efficacy and safety of the drug product. As one of the global leaders in protein science and formulation science, CD Formulation offers a wide range of forced degradation study tests to help you understand how your product performs under extreme conditions and supports different development stages, including early development phases and late development phases.

Purpose of Forced Degradation Studies

Forced degradation studies involve intentionally breaking down protein/peptide molecules to an appropriate degree through various stresses such as pH, temperature, light, chemical agents (e.g., oxidants, deoxidants, etc.), and mechanical stresses to accelerate the chemical degradation, physical degradation, and instability of these biopharmaceuticals. Forced degradation studies can provide a range of information about the possible degradation products of a particular biopharmaceutical. This information helps determine the degradation pathways and intrinsic stability of the molecule and is used for a variety of purposes, including:

• Establish degradation pathways for protein/peptide drug products.
• Identify possible degradation products.
• Determine the intrinsic stability of protein/peptide in the formulation.
• Distinguish between drug-related degradation products and non-drug degradation products in formulation.
• Reveal possible degradation mechanisms of protein/peptide, including hydrolysis, oxidation, thermal, or photolysis.
• Establish stability-indicating properties of developed methods.
• Understand the chemical properties of protein/peptide drug molecules.
• Support formulation development.
• Support process development.
• Support comparability or similarity studies.
• Support formulation stability and batch release studies.
• Resolve stability-related issues.

Fig. 1 Purpose of forced degradation studies for protein/peptide drug products. (CD Formulation).

Explore Our Proteins & Peptides Forced Degradation Studies

Regulatory agencies require thorough studies of the stability of drug substances and their degradation under stress conditions to ensure the quality, safety, and efficacy of biologics.

At CD Formulation, our analytical team can help you conduct comprehensive and thorough forced degradation studies to obtain meaningful data on degradation mechanisms, including biochemical and biophysical properties of the drug, its main degradation pathways, degradation protein forms, and unstable sites, and identify product-related variants.

In addition, this data can help you understand how your product behaves under extreme conditions, such as during transportation, packaging, etc.

Throughout the forced degradation study process, our team of scientists subject the protein/peptide drug or final product to a range of stress conditions.

Typical Stresses Include:

• High temperature.
• Freeze-thaw cycles.
• Mechanical stress, including shaking, stirring, and rotating.
• Photostability testing according to ICH.
• pH stress, including acid, and base.
• Forced oxidation.
• Forced deamidation.
• Shipping studies.
• In-use stability studies.
• Compatibility studies with various materials and drug delivery systems.

Our Expertise in Forced Degradation Studies

Protein/peptide biologics can often degrade in many different pathways with varying kinetics. The magnitude of the forced degradation conditions needs to provide measurable changes and identify the most relevant degradation pathways. Our team of scientists designs conditions based on known and expected degradation pathways and experience with similar molecules to identify possible degradation pathways, including oxidation, hydrolysis, photolysis, and aggregation.

Oxidation

Oxidizing conditions are primarily created by exposure to atmospheric oxygen, through the action of light, heat, moisture, and agitation. The side chains of methionine, cysteine, histidine, tryptophan or tyrosine residues are susceptible to oxidation, with methionine being the most reactive residue. We offer oxidation conditions:

Hydrolysis

Hydrolysis is the breaking of peptide bonds between amino acid residues, releasing smaller peptide chains. Hydrolysis is mainly caused by exposure to acidic or alkaline pH. We provide hydrolysis conditions:

Photolysis

A range of photolytic reactions occur upon exposure to light, including aggregation or peptide bond cleavage.

Aggregation

Aggregation is often a result of mechanical stress (e.g. shaking, stirring, rotation) as well as oxidation, hydrolysis, and photolysis as mentioned above. In addition, intramolecular chemical bonding, such as the reaction of amino acid residues with trace metals (copper or iron) or incomplete protein reduction, can also produce aggregation. We are able to perform thorough characterization to assess the presence of insoluble aggregates and identify possible degradation products.

Forced Degradation Studies During Development Stage

Forced degradation studies can be performed in early or late development, depending on the purpose of the study and the amount of material available. CD Formulation is able to support forced degradation studies at any stage of your product development to provide knowledge about the degradation pathways of your molecule.

Early Development Stage

Our forced degradation studies can help you develop optimal processes and formulations. In addition, degradation samples can help develop stability-indicating analytical methods by demonstrating whether current methods are adequate to assess stability (e.g., using oxidized samples to develop methods to determine oxidized form) and by determining which test parameters are the best indicators of stability.

Mid/Late Development Stage

Limited materials early in development limit the completeness of forced degradation studies, and process steps and formulations may also change during later development. If needed, we also provide more comprehensive forced degradation studies later in development to support the development of your protein/peptide drug product in preparation for commercial launch.

Our Technology Platforms

Given the complexity of protein/peptide biopharmaceuticals, there is no single stability-indicating method that can describe all of their stability characteristics. Our protein scientists deploy a range of analytical strategies to conduct forced degradation studies to determine identity, purity, content, and bioactivity, and monitor impurities.

• Appearance (i.e., color, clarity, particulate matter): Optical microscopy, laser particle size analyzer.
• Physical and chemical properties: Differential scanning calorimetry (DSC), circular dichroism (CD).
• Aggregates: Size exclusion high-performance liquid chromatography (SE-HPLC).
• Protein content: Amino acid quantification, reversed-phase high-performance liquid chromatography (RP-HPLC), UV-visible spectrophotometry.
• Deamidation: Isoelectric focusing (IEF)/imaging capillary, isoelectric focusing/ion-exchange high-performance liquid chromatography (IE-HPLC).
• Purity and specific impurities: Reversed-phase high-performance liquid chromatography (RP-HPLC).
• Protein structure identification: CD, Fourier transform infrared (FTIR), x-ray crystallography, nuclear magnetic resonance (NMR), cryo-electron microscopy (cryo-EM), hydrogen-deuterium exchange (HDX)-MS, and small-angle x-ray scattering (SAXS).
• Bioactivity assays: In vitro binding assays such as enzyme-linked immunosorbent assay (ELISA)and surface plasmon resonance (SPR), and functional bioassays such as in vitro cell proliferation and in vivo animal models.

Why Choose Us for Proteins & Peptides Forced Degradation Studies？

• Our team of scientists has extensive experience in conducting forced degradation studies for proteins and peptides, ensuring accurate and reliable results.
• We adhere to strict quality-control measures and follow regulatory guidelines for forced degradation studies, guaranteeing high quality data and analysis.
• Our laboratory is equipped with advanced instrumentation and technology to carry out comprehensive forced degradation studies on proteins and peptides.
• We work closely with clients to develop a customized study plan that meets their specific needs and requirements, ensuring the most relevant data is obtained.
• We provide flexible test options and customized solutions to meet your needs at different research stages.

Publication

CD Formulation has decades of experience in using forced degradation studies to support biopharmaceutical development. We consistently provide total quality assurance expertise to help you ensure your quality, safety, and efficacy requirements are optimally met and exceeded. Please feel free to contact us if you are interested in our services or have further questions. We look forward to working with you to promote development and innovation in the biopharmaceutical field.

How It Works

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

STEP 2

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STEP 3

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