Better, Faster, Stronger: 5 Trends Impacting Biopharmaceutical Science

While 2020 has been an incredibly trying year, it has also resulted in notable achievements. These milestones, from vaccine development to new modalities, have transformed the biopharmaceutical industry. Here, we look back on five trends that have influenced the industry this year that will continue to shape it in 2021 and beyond.

More analytes than ever before
Traditional engineered protein and monoclonal antibody drugs continue to account for a large percentage of new biologics under development, but next-generation modalities including cell and gene therapies, multi-specifics and genetic vaccines and therapeutics, to name just a few, are experiencing explosive growth. 

Consequently, there is an increasing need for highly flexible analytical systems useful for the rapid characterization of many types of analytes with sensitivity, precision and high resolution. These systems must be capable of separating, detecting and identifying multiple analytes simultaneously across a broad range of concentrations in complex matrices.

Often the analytes have highly similar structures, such as nanobodies differentiated by just a deamidation or two. Protein contaminants from host cells and media, which can have a negative impact on safety and efficacy, also present analytical challenges. Developing an effective single assay that can analyze the different host-cell protein (HCP) profiles (up to 1,000 or more in each host cell) has not been possible using conventional ligand binding assays.

A simple, quick, and effective technique with exceptional resolving power and a high degree of precision, capillary electrophoresis (CE) is increasingly being used to check and confirm the purity, heterogeneity, and glycan association of biologic drugs of all types. Specialized and standardized reagents and kits optimized for specific CE methods, such as capillary isoelectric focusing (CIEF), capillary zone electrophoresis (CZE), CE-sodium dodecyl sulfate (CE-SDS) and purposes CE – laser induced fluorescence (CE-LIF) offer complete workflow solutions that are precise, but also simple and flexible enough for quality control applications.

CE is also being combined with mass spectrometric methods for a range of analyses required during the development and commercialization of next-generation modalities. Meanwhile, liquid chromatography tandem mass spectrometry (LC-MS/MS) leveraging a data-independent acquisition technique has proven to offer substantially more comprehensive coverage, dramatically faster and simpler assay development, better indications of biotransformation and analyte characterization, fewer false negatives, and cheaper reagents when multiplexed. For instance, this method can detect HCPs from different organisms simultaneously and identify and quantify all the HCPs in a single injection regardless of their concentration. In addition, it can be applied to any biologic, including cell and gene therapies, without the typical 1.5-2-year development effort.

Demand for novel vaccines at all-time high
The COVID-19 pandemic has spurred unprecedented activity in the development of new vaccines. A range of traditional and cutting-edge vaccine development approaches are being pursued. Genetic vaccines based on naked DNA plasmids, viral vectors and mRNA that leverage robust, scalable platform manufacturing concepts and integrated processes have shortened timelines. Advanced analytics are playing a crucial role in ensuring the safety and efficacy of these new vaccines as they are rapidly developed and commercialized.

Many analytical technologies initially developed for protein therapeutics have been modified to address the assessment needs for genetic vaccines. CE-LIF provides a rapid, sensitive, reproducible, and automated method for the quantitative analysis of plasmid DNA isoforms and mRNA detection, separation, and sizing. It is important to consider what unique chemistries to analyze when completing routine or difficult automated Sanger genome sequencing.

LC-MS/MS can be leveraged for identification and quantification of HCPs and other contaminants in viral vector capsids. Confirmation of the LNP composition of formulated mRNA vaccines can also be achieved using LC-MS/MS with a targeted lipidomic assay, scheduled multi-reaction monitoring (MRM) and fast polarity switching, allowing the identification and quantification of nearly 1150 different polar and neutral lipids.

Regardless of the vaccine technology, confirmation that the desired antigen is expressed, and adequate T-cell responses are achieved, is essential. State-of-the-art MS, CE-LIF and multiplex genome analyses can help support manufacturers in this confirmation phase.

Unchartered territory with new modalities
In addition to new genetic vaccines, many new RNA- and DNA-based therapies such as oligonucleotide antiviral drugs, viral and other gene therapies, various types of cell and gene-modified cell therapies, bi- and tri-specific antibodies, antibody-drug conjugates bi-specific T-cell engagers, peptibodies and nanobodies are under development today.

These newer modalities overcome some of the limitations of monoclonal antibodies (mAbs), such as enabling simultaneous binding to multiple sites, greater stability, and the ability to access solid tissues and cross the blood-brain barrier. However, the complexity of these new modalities and processes can yield numerous variants. Titers are also generally much lower (10-50%) than those for mAbs.

The diversity and greater complexity create a higher analytical burden beginning at the clone selection stage through process development and commercial production.  It is necessary to distinguish molecules with minor structural differences at low concentrations. Additional sensitivity and separation resolution are therefore essential when developing analytical methods for these new modalities.

To overcome these challenges, existing trusted mAb methods are being adjusted for variants during assay development. As an example, CE-SDS, cIEF, CZE and fast glycan analyses for peptibody and nanobody analyses may be optimized by increasing the percentage of reagents in the sample, using different reagents, lowering the pH and changing the temperature and time of the analysis.

The industry has been working towards alternative orthogonal techniques (instead of just modified mAb methods) that can address the specific complexity of mAb variants. Hyphenated techniques such as Capillary Electrophoresis paired with mass spectrometry (CE-MS) can support analysis of charge variants of intact nanobodies even with mass differences of just 1-2 Daltons.

High resolution mass spectrometry (HRMS) can ensure sufficient resolution of parent oligos and major and minor metabolites for oligonucleotide antiviral drugs.

For characterization of multispecifics on the subunit level, increased throughput can be achieved using a LC-MS/MS system with differential mobility separation (DMS) technology. This technique enables separation of protein subunits and unambiguous characterization of each chain with a single injection and without the need for chromatographic separation, reducing the overall time required to complete studies.

Patients need therapies quicker
For developers of both traditional and next-generation therapies, time to market is critical. For new modalities that target specific genes, the urgency is even greater; the first to market wins ─ there is no second place.

The key to improving the manufacturing of gene and other novel therapies is therefore development of consistent, scalable, high-yielding platform processes and rapid analytical methods. Without speedy assays, the ability to fully understand all the relevant process parameters and how they impact product quality attributes is limited, which prevents the development of robust processes.

Advances in automation and data analysis have the potential to reduce analysis times as well as simplify analyses while increasing consistency and accuracy. These assays must also have greater sensitivity and precision given the often-low production volumes for next-generation therapies.

CE solutions have been shown to offer the high-sensitivity and high-resolution required for GMP release for various applications. AAV capsid protein purity, for instance, can be determined using CE-LIF with sensitivity four orders of magnitude greater than traditional SDS-polyacrylamide gel electrophoresis (PAGE) method and a much higher throughput using a smaller sample quantity.

Need for analytical maturity
The need to develop established, qualified processes and analytics sets new modalities apart from monoclonal antibody drugs, which have clearly defined process and analytical requirements. Lacking platform solutions, regulatory guidelines and often skilled and experienced personnel, developers of gene, cell and other next-generation therapies must create their own paths to commercialization.

The US Food and Drug Administration (FDA) does, however, continue to develop guidance documents to support the development and commercialization of these novel, life-changing medicines. FDA encourages advances by creating opportunities for discussing the best approaches to ensure the safest products.  Various consortia of technology providers, drug developers and regulatory agencies are working together to modify existing methods and developing new techniques that simplify and reduce the time required for analysis of novel modalities.  Coupling the best analytical technologies with the brightest ideas in treating diseases will bring novel therapies to patients as rapidly as possible.