Good Manufacturing Practice (GMP) stability testing and GMP release testing for large-molecule biopharmaceuticals and proteinaceous In Vitro Diagnostic (IVD) reagents are challenging due to the complex nature of these products and the stringent requirements for their use in clinical settings. Large-molecule biopharmaceuticals, such as proteins, antibodies, and nucleic acids, are structurally complex and often sensitive to environmental conditions, which makes it difficult to maintain their integrity over time and during testing. Because this structural complexity requires sophisticated analytical techniques to fully characterize each molecule and ensure batch-to-batch consistency, partnering with a bioanalytical CRO that offers state-of-the-art bioanalytical services is essential.

The assessment of large-molecule biopharmaceuticals typically involves advanced and precise analytical methods that must be meticulously developed, validated, and standardized to ensure they are sensitive enough to detect impurities and degradation products, as well as confirm molecular integrity. Determining the biological activity of large molecules is more complex than measuring the concentration of small-molecule drugs. Bioassays, often used for this purpose, are inherently variable and require live cells or other biological systems. Ensuring the reproducibility and accuracy of these bioassays is a significant challenge, and it is essential to work with a bioanalytical CRO with proven bioanalytical laboratory services capable of handling these demands.

GMP Stability Testing Challenges for Large-Molecule Drugs

The purpose of GMP stability testing is to understand how the quality of an active pharmaceutical ingredient (API) varies when different environmental factors, such as temperature, humidity, and light, are introduced. GMP stability testing also includes the study of product-related factors that influence the quality of a drug, such as the interaction of API with excipients, container closure systems, and packaging materials. GMP stability testing informs a drug product’s shelf life, suggests appropriate storage conditions before and after opening, and identifies potential degradation or quality concerns. Stability assessments are performed in nearly every development phase to evaluate a drug product’s overall stability in various instances.

Large-molecule drugs face unique GMP stability testing challenges. Their greater complexity, combined with more batch-to-batch variability and specialized storage conditions, makes typical stability study conditions unsuitable. This means bespoke stability testing programs must be designed for each product. Biopharmaceuticals are generally highly concentrated and/or less soluble, which increases the likelihood that the API will precipitate during stability studies. Clear, quality data surrounding this process are essential for ensuring regulatory compliance, underscoring the importance of partnering with the right bioanalytical CRO.

Degradation pathways of biopharmaceuticals are much more unpredictable than for small-molecule drugs, and they vary for different proteins. For example, some parenteral biologics administered to patients are highly concentrated and may precipitate during the stability studies.

Biopharmaceuticals are often only stable over a very limited temperature range, meaning that temperature deviations outside the optimum storage conditions can significantly impact stability. This means they must be stored within an extremely narrow temperature range to avoid an impact on biological activity. GMP stability testing of such sensitive products in a range of temperatures must be carefully planned to consider actual storage conditions. The stability of proteins often also calls for analytical methods other than liquid chromatography, which is frequently used for small organic molecules.

GMP Release Testing Challenges

GMP release testing is a critical quality control process involving physical, chemical, and microbiological tests. These tests ensure that manufactured therapeutics meet predetermined specifications before being released to the market. GMP lot release testing must demonstrate that products released to the market are consistent with those assessed during the regulatory process.

The importance of GMP lot release testing lies in its ability to verify that manufacturing processes are operating correctly and consistently, ensuring the quality and safety of the products.

The amount of testing required for each batch depends on various factors, such as identified risks, variability in the manufacturing process, and the testing methods applied. A sampling procedure should be established to ensure that samples taken for analysis are representative of the manufactured batch.

Appropriate control measures should be built into the manufacturing processes in the early phases of product and process development to minimize the need for extensive GMP lot release testing. Regulators typically require companies to provide suitable batch records, including evidence of the batch testing that has been performed. These records should follow the ALCOA standard (Attributable, Legible, Contemporaneous, Original, and Accurate) and provide full traceability of the manufacturing process. A reputable bioanalytical CRO will take the appropriate measures to ensure regulatory compliance.

Prolytix GMP Stability Testing and Storage Services

Prolytix leverages 35+ years of protein chemistry experience to support the large-molecule drug product lifecycle from discovery through release, including implementing and performing drug stability testing procedures for large-molecule biopharmaceuticals and proteinaceous IVD reagents. The bioanalytical CRO offers real-time and accelerated stability programs and shelf-life determination. Prolytix performs all drug stability testing under strict GMP compliance in its full-service, FDA-inspected GMP facility. Therapeutic developers know exactly how their product performs under a wide range of conditions—all of which meet ICH guidelines and the compliance requirements of leading global regulators. Prolytix provides testing services for clinical trial materials and products to prepare for commercial release.

Prolytix uses client-defined, compendial, or in-house developed and validated methods to provide compliant stability testing data. Prolytix also offers ICH-compliant stability storage on-site.

Prolytix GMP Storage Services

Prolytix helps control overhead costs and maintain product integrity with reliable stability storage services for clinical and commercial materials. The bioanalytical CRO’s GMP storage facility provides ICH-compliant storage under a wide range of temperature and humidity requirements. Backup storage and on-site generators help safeguard materials in the unlikely event of a chamber or power failure.

Prolytix GMP Release Testing Services

Prolytix provides GMP release testing for commercial and clinical trial drug substances and drug products. As a bioanalytical CRO, we test bulk, final product, and in-process materials in its fully compliant, FDA-inspected facility. Combined with Prolytix’s other Chemistry, Manufacturing, and Control (CMC) and drug development services, biotherapeutic developers are assured the highest quality from early-phase clinical development through commercial release.

• Prolytix performs validation studies on all tests and processes to demonstrate product compliance.
• Prolytix conducts strict sample reconciliation against the documentation provided. After set-down, Prolytix’s environmental monitoring system oversees all storage chambers that house samples. Backup storage and generators are on-site in the unlikely event of a chamber or power failure.

GMP Release Testing Analytical Methods

Prolytix helps develop analytical methodology and acceptance criteria, as well as execute predefined GMP release testing. Prolytix emphasizes customer success and provides fast, reliable turnaround and efficient sample management.

• Compendial Methods
• Osmolality Measurement
• Residual Moisture Determination
• Excipients including pH Modifiers, Tonicity, Amino Acids, Preservatives, Chelators, Surfactants, Inhibitors
• HIAC Liquid Particle Analysis
• Thrombin Generating Assays
• HPLC/UPLC Methods (Reverse Phase, Size Exclusion, Ion Exchange)
• SDS-PAGE
• Clotting Assays (Factor Assays, PT, aPTT, NAPTT)
• Anti-Drug Antibody Assays
• Relative Potency
• Protein Concentration