Analytical skills are needed in biological product testing

Personnel should be trained in various sets of analytical skills for testing biological drug substances.

The complexity of biological drug substances requires specialized analyzes and the training necessary to perform the analyses. It is not enough to know how to run the tests, but also how to interpret the data to give meaningful clinical value to the analyses. To better understand the methods by which the results of biological drug substance testing should be analyzed and interpreted as well as to better understand the challenges inherent in the treatment of complex biological molecules, Pharmaceutical technology spoke with Khanh Ngo Courtney, Senior Director of Biologics at Element, and Mahesh Bhalgat, COO of Syngene International.

The need for skills

Pharma Tech: Is there currently a shortage of laboratory personnel with the specific skills to interpret data from biological drug substance testing? If so, what vocational training is most needed?

Courtney (Element): Increasingly, we find that the characterization and routine testing of biological therapeutics, especially advanced therapeutics such as gene and cell therapies, requires advanced analytical techniques combined with a deep understanding of biology, which requires a specialized laboratory staff. For example, enzyme-linked immunosorbent assay (ELISA) is becoming a less efficient way to quantify host cell protein (HCP) clearance due to emerging data showing that antibodies generated against HCPs do not provide sufficient protein coverage. of the host cell. Therefore, a more comprehensive analysis of health professionals is needed. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) appears to be a necessary analytical method to quantify and characterize the clearance of PHCs. It is difficult to find talent who can develop and run a robust quantitative LC-MS/MS method suitable for Chemistry, Manufacturing and Controls (CMC).

Similarly, biological therapy requires a in vitro cell assay to show potency during characterization and for release and stability testing. The design and development of the appropriate cell power methods, including the readout method, requires personnel who understand the mechanism of action of the therapeutics and how to exploit the cell biology toolkit to show in vitro drug efficacy in a robust and CMC-appropriate manner. The appropriate skills required to perform this type of work are not easily found in one person.

Bhalgat (Syngene): Biologic drug testing is changing rapidly and new platforms and technologies are being used for testing purposes to meet the needs of drug testing and drug mechanisms. While many laboratories and laboratory personnel are technically sound, there is still a lack of understanding and decision-making on the use of analytical models for the characterization of biologicals and for studies of the mechanism of biologics. share (MoA). We find that the greatest shortage of laboratory personnel is for scientists trained in statistical designs, analysis and validation of qualitative tests.

Additionally, there is a gap in recruiting professionals with training in biological analysis, particularly in the areas of structural and functional characterization. Besides the physicochemical field, there is a need to strengthen the knowledge base on analytical ion exchange chromatography and N-glycan analysis for glycosylated biologicals.

Interpret/analyze data

Pharma Tech: What have been some of the key challenges in interpreting and analyzing data generated from drug substance testing on biological APIs?

Bhalgat (Syngene): Drugs, including APIs and biologics, must be manufactured to appropriate quality standards. Some of the major opportunities associated with the analysis and interpretation of data from biologics testing include the identification and analysis of product and process related impurities, which is always challenging due to the specialized nature tests ; assay changes during stability studies due to interaction-induced non-covalent oligomer formation; and HCP analysis of samples being processed.

The main challenge lies in interpreting the data for its significance when missing large datasets generated over an extended period of time. Because data trend monitoring and analysis is important for biologics and other drugs, we apply quality-by-design (QBD) approaches to ensure data interpretation and comparison needs are met. .

Courtney (Element): The heterogeneity of the biological molecule has an impact on the way methods are developed, data are generated and interpreted. Large molecules are tens, if not hundreds, of thousands of small molecules that chemically interact with each other to form tertiary structures, which translate into function. Yet not all molecules in a solution of the drug substance will be exactly the same – they vary in the amount of post-translational modifications such as glycosylation, phosphorylation and other chemical changes as well as the damage they undergo, such than amino acids. deamidation, reduction and acid oxidation. All of these chemical changes impact the structure of the biological molecule, its function, and how it interacts with other molecules in the cell and how it behaves in analytical methodologies.

Pharma Tech: What methods or approaches have so far been successful in generating clinically meaningful data and analyzes of that data?

Courtney (Element): Meaningful data in the clinic require the appropriate biomarkers to be selected, and the methods for detecting and quantifying said biomarkers are robust and sensitive. Meso Scale Discovery (MSD) and Quanterix immunological technologies are common methodologies and have proven successful in the analysis of clinical samples. LC-MS/MS, if available, remains a powerful tool for clinical data analysis for its sensitivity and robustness in detecting and quantifying not only large molecules, but also small molecule biomarkers.

Bhalgat (Syngene): In clinical research, obtaining meaningful data depends on several factors. Best practices that have been successful in generating clinically meaningful data and that have facilitated the interpretation/analysis of the generated data include drug-specific pharmacokinetic (PK), pharmacodynamic (PD), and immunogenicity tests, rather than a generic/ready to use kit assays or methods. Validated methods are also necessary to ensure the performance of the test, in accordance with expectations regarding the defined dosage, route of administration and planned clinical interventions. Additionally, careful review of the clinical trial protocol early in trial development, including a two-way dialogue between the clinician and the bioanalytical scientist, helps to establish methods that are appropriate for the particular trial. Finally, paying close attention to the performance of the method for a particular population of test subjects, their age, disease state, ethnicity, etc., helps generate reliable data.

Respond to industry needs

Pharma Tech: Where is the most unmet need for analytical solutions for complex and difficult biological drug substance testing?

Bhalgat (Syngene): Bioanalysis of biologics requires the ability to use many different approaches, so there are many unmet needs. For example, there is a need for high-throughput automation and convenient biostatistics for complex and challenging biological drug substance testing. Meanwhile, immunoassays for biologics are labor intensive and people dependent. Moving to much more automated technology to perform sensitive and specific immunoassays is a clear opportunity. There is also a need for assays for biophysical characterization in the presence of various process matrices, host-specific, HCP assays, and non-destructive assays to understand the tertiary structure of drugs at the atomic level (e.g., cryo-microscopy electronic or monomolecular fluorescence techniques are not yet available for industrial use). Additionally, due to the lack of proper guidelines, there are regulatory differences between different geographies, resulting in the development of varying approaches to product testing.

Courtney (Element): Highly complex biological drug substances, such as genetic information in the form of viral therapeutic agents or encapsulated in nanoparticles (eg, lentivirus, adeno-associated virus [AAV], nanoparticles), are extremely difficult to analyze due to heterogeneity in drug substance quality. An example is the analytical challenge of separating and quantifying viral particle fill amounts (i.e., empty vs. partial vs. full). When lentiviral AAVs or APIs are produced, part of the drug substance solution will include viral particles containing the correct number of copies of genetic information, particles containing too much material, particles with too little, and particles containing no equipment. Having the analytical capability and technology to achieve resolution between these different drug substance molecular compositions is still an unmet need. Current methods all have their own pitfalls. Analytical ultracentrifugation is expensive, not very robust and requires a large amount of sample. Transmission electron microscopy is time-consuming and lacks quantitative power. High performance liquid chromatography has low resolution. Finding the right analytical solution for the intended purpose is an unmet need for many methods defining the quality of advanced therapeutic molecules today.

About the Author

Feliza Mirasol is the scientific editor of Pharmaceutical technology.

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