Uncover and Eliminate Process-Related Impurities in Biologics
Various chemicals are introduced throughout the pharmaceutical and biologic drug manufacturing process to enhance product expression and recovery. While most of these chemicals are eliminated during downstream purification processes, lingering impurities can affect the safety and efficacy of your final product, delaying drug development and market release. Collaborate with our team of specialists, who employ highly sensitive impurity testing techniques tailored to accurately define the limit of detection (LOD) and limit of quantitation (LOQ) using your samples and matrices. Prior to using assays for CGMP lot release programs, we follow all required qualification and/or ICH validation guidelines.
| Residual | Method Description | LOQ (ppm)1 | LOD (ppm)1 |
|---|---|---|---|
| IPTG | HPLC with ELSD/CAD | 6 | < 1 |
| PEI | HPLC with ELSD/Fluorescence | 10/< 2 | 1-2/< 0.5 |
| Kanamycin | HPLC with UV/Vis | 0.03 | < 0.01 |
| Anti-foam | HPLC with ELSD/CAD | 1 | 0.1 |
| TFA/Acetate | HPLC with UV/Vis | 1 | 0.1 |
| Cyanate | HPLC with Conductivity/ECD | 0.6 | 0.1 |
| Imidazole | HPLC with Fluorescence | 0.13 | < 0.1 |
| Triton X-100 | HPLC with ELSD/CAD | 5 | 1 |
| Polysorbate (tween) 20/80 | HPLC with ELSD/CAD | 5 | 1 |
| Zwittergent | HPLC with ELSD/CAD | 1-5 | < 1 |
| Pluronic | HPLC with ELSD/CAD | 1-5 | < 1 |
| Protein A | ELISA | 0.001-0.10 | < 0.001 |
| PEG | HPLC with ELSD/CAD | 1-10 | < 0.5 |
| TRIS | HPLC | 1-10 | < 0.5 |
| Growth Factors (e.g., insulin, interferon) | ELISA | 0.001-0.10 | < 0.001 |
Product-Related Impurities in Biologics
Along with process residuals, there is also the possibility for product-related impurities to be carried through the manufacturing and purification process. These impurities, including host cell proteins, host cell DNA, N-terminal truncations, and other potential modifications, may cause adverse reactions in animals and humans, and therefore, impurity testing must be employed to ensure that any contaminants present are below pre-determined acceptable levels.
Our comprehensive impurity testing solutions are designed to detect and quantify a broad spectrum of potentially harmful compounds from different biological mediums, increasing your product’s quality and safety. From host cell proteins to host cell DNA and N-terminal truncations, our dedicated team of specialists will leverage state-of-the-art technologies to help you deliver safe and effective treatments for those who need them most.
| Impurity | Impurity Description | Method Description | LOQ (ppm) | LOD (ppm) |
|---|---|---|---|---|
| Host cell proteins (HCP) | HCPs are unwanted impurities released during biopharmaceutical production, specifically during cell culture. Samples should be carefully scrutinized for HCPs to maximize the safety of your therapeutic. | Commercial kit or customized assay | 0.001-0.010 | < 0.001 |
| N-terminal truncations | During biopharmaceutical production, N-terminal truncations may arise and must be identified to ensure the biological activity, stability, and safety of the resulting proteins. | N-terminal sequencing | > 10% | |
| Host cell DNA | Cells used to produce biopharmaceuticals such as recombinant proteins may release unwanted DNA fragments. These should be identified to mitigate the risk of adverse immunogenic reactions. | PCR | Method- and product-dependent | |
| Oxidation/deamidation (PTMs) | PTMs such as oxidation and deamination may occur during protein-based pharmaceutical manufacturing. Like N-terminal truncations, these modifications must be monitored, as they can impact your biologics' stability, efficacy, and safety. | Peptide mapping LC-MS or LC-MS/MS, IsoQuant HPLC | Method- and product-dependent | |
| Isoform analysis | Isoform analysis identifies and quantifies the various forms of a protein that arise from genetic variation and modification. Eliminating unwanted isoforms is critical to free your pharmaceuticals of unintended variants that may affect their safety. | Ion exchange HPLC, cIEF, sialic acid analysis | Method- and product-dependent | |
| Disulfide bridge scrambling (mis-folding); conformational impurities | Disulfide bridge scramblings arise when a protein’s cysteine residues mispair, causing misfoldings and altered structure and function. This can significantly impact the efficacy and immunogenicity of your pharmaceutical. | Peptide mapping LC-MS or LC-MS/MS, Ellman’s free sulfhydryl, intrinsic tryptophanfluorescence, ANS dye binding, NMR | Method- and product-dependent | |
| C-terminal integrity | The C-terminal integrity of your therapeutic proteins should be carefully scrutinized for truncations and alterations to mitigate instability, inactivity, and overall dysfunction. | Peptide mapping LC-MS or LC-MS/MS | Method- and product-dependent | |
| Aggregation | During biopharmaceutical manufacturing, some proteins are susceptible to aggregation, resulting in dimers and oligomers with reduced therapeutic activity. Detecting these aggregations is vital to ensure the proper function, stability, and safety of your biologic. | Analytical ultracentrifugation (AUC), SEC/SEC-MALLS, dynamic light scattering (DLS) | Method- and product-dependent | |
Frequently Asked Questions (FAQs) About Impurity Testing
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What is the difference between impurity testing and residual testing?
Both impurity and residual testing are vital to ensure the safety and efficacy of pharmaceuticals. Residual testing detects and quantifies substances that are intentionally incorporated during the manufacturing process but are unwanted in the final product. Impurity testing, on the other hand, identifies compounds and byproducts that are introduced or formed during the manufacturing process or as a result of degradation over time. Both of these tests are vital for regulatory compliance and ensure the quality and safety of pharmaceuticals.
Collaborate with our dedicated team of scientists and benefit from our expertly operated technologies and methods. Together, we’ll screen your biopharmaceuticals for a comprehensive spectrum of impurities with precision and reliability, ensuring your product’s quality and safety.
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What technologies and methods are used in impurity and residual testing?
The methods and technologies used to identify and quantify impurities and residuals are selected based on the nature of the compounds being tested, the matrix of the sample, and the level of desired sensitivity.
High-performance liquid chromatography (HPLC) is versatile and widely used to separate, identify, and quantify impurities and residuals with high sensitivity and accuracy. For compounds not easily detected by UV/Vis absorbance, HPLC can be coupled with evaporative light scattering detector (ELSD) and charged aerosol detector (CAD) technologies to expand HPLC’s application range in impurity and residual testing. Mass spectrometry can also be implemented to complement HPLC for high specificity and sensitivity testing. With the ability to identify unknown impurities, mass spectrometry can be an effective tool to gain valuable insights into the formation of unwanted residuals during the manufacturing process.
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What is the difference between product-related and process-related impurities?
Both product and process-related impurities are released throughout the pharmaceutical manufacturing process. Product-related impurities are directly associated with the drug substance itself and may be byproducts or degradation products formed during the synthesis, storage, and handling of pharmaceutical ingredients. In addition, they may result from underlying chemical reactions such as oxidation, hydrolysis, and photodegradation. Process-related impurities, on the other hand, originate from the raw materials, intermediates, and reagents used in the biomanufacturing process. With impurity testing, valuable insights can be gained to optimize the manufacturing procedure, resulting in fewer product and process-related impurities.