SPECIAL FEATURE – Analytical Testing Trends in 2022
An increasing number of clinical trial registrations, more R&D investment, growing demand for biopharma products, a continued focus on safety and quality, and more third-party testers entering the market are key reasons why the US pharmaceutical analytical testing outsourcing market is expected to reach $5.55 billion in the next five years.1 Globally, the market could reach $12.4 billion by 2028.2
Specifically, bioanalytical testing is anticipated to experience the fastest growth over this period.1 In fact, this segment led the global market in 2020 due to the high number of clinical trial registrations.2
This annual Drug Development & Delivery report reveals the innovative technologies and techniques that leading outsourcing providers currently offer for both small and large molecules.
Alcami offers comprehensive services to support biologic drug product development from preclinical/early-phase programs through commercial. Its bioanalytical testing services are designed to support drug substance and drug product development, validation, and routine analysis, including associated raw materials, excipients, components, and finished goods. Typical programs include therapeutic proteins, peptides, and nucleic acid products. Capabilities consist of cell-based assays with associated cell culture workflows, ELISA, electrophoresis, amino acid analysis, HPLC/UPLC with various detection modes such as UV/PDA, CAD, ELSD, RI, FLD, and MS.
“Alcami utilizes high resolution MS for intact mass analysis. We perform identity testing of post-translational modifications, glycan analysis, and peptide mapping,” explains Katie Schlipp, Vice President, Laboratory Operations, Alcami. “Additionally, these products are commonly analyzed by ion pairing chromatography, ion-exchange chromatography, and PCR-based approaches.”
Alcami also offers a high-throughput method development for efficient screening of solvents and columns. This platform allows Alcami to select optimal parameters quickly and develop robust and QC-friendly methods in much shorter timeframes, says Ms. Schlipp.
There are some specific key market trends that Alcami is seeing in the industry. One is the need for microbial in-use or admixture compatibility studies. A microbial in-use study is intended to evaluate the growth of a low level of microorganisms inoculated into the diluted product over the hold time to represent inadvertent contamination during rehydration or dilution of the product.
These studies determine if the product will support the growth and/or proliferation of this inadvertent contamination during the holding period prior to patient administration. “Our team of experts can support the experimental design and execution of in-use studies and provide a final scientific report for our client’s filing,” she says.
A second trend is an increase in the need for environmental monitoring. “The global health crisis triggered by the COVID-19 pandemic has increased the demand for sterilized pharmaceutical formulations and scientific advancements in cleanroom technology,” says Ms. Schlipp. “Alcami experts can guide customers through the stringent regulatory framework associated with cleanroom space and provide support specifically tailored to the needs of each cleanroom to ensure that quality, safety, and efficacy are maintained.”
She adds that a successful environmental monitoring program begins with the appropriate risk assessment, qualification, and certification activities for the space and utilities that lay the groundwork for routine testing. To this end, Alcami provides support in personnel plating, sampling performed during client manufacturing activities, and expertise in remediation efforts in case of unfortunate events such as failure to HVAC systems.
She says: “Environmental services are holistically enhanced by Alcami’s microbiological services, from microbial and fungal identification to disinfection efficacy and biological indicators testing.”
Beyond these capabilities, Alcami is investing in new capabilities, including analytical ultracentrifugation, variable pathlength UV for A260/A280 determinations, and new Tandem Quadrupole and QTOF instruments as part of a 16,000-square foot renovation in its Durham, NC, laboratory to be commissioned early this year.
Along with the significant investments in analytical capabilities and expansions, Alcami recently announced the acquisition of Masy BioServices. “GMP storage is a critical need in the pharma industry,” says Ms. Schlipp. “Masy offers secure and tightly controlled GMP temperature storage from -196°C to 70°C, including all ICH stability conditions, for various materials including vaccines, biopharmaceuticals, cell banks, tissues, compounds, and medical devices. In addition, pharma support services through Masy include equipment calibration, large-scale validation and qualification projects, SenseAnywhere monitoring system, and equipment sales, and rentals.”
Analytical testing is a crucial part of the drug development process for both small and large molecules. And, Muhammad Asif, PhD, executive director, Analytical R&D and Quality Control, Ascendia Pharmaceuticals, says that Ascendia’s analytical testing has rescued important drug development programs. For example, Ascendia has developed methods that have predicted and helped control the in vivo time-dependent release of an injectable drug that precipitates at the injection site and releases slowly. “Such site-specific injections are increasingly becoming common and are made in a specific organ, joint or intervertebral space,” Dr. Asif says. “These drugs are intentionally designed to precipitate or coagulate to provide maximum effect at the inject site without creating unwanted toxicity by lowering systemic absorption.”
Whether testing liposomal drugs or drugs embedded in a polymer, Ascendia has methods to analyze drug content and release. High-speed chromatography such as Ultra Performance Liquid Chromatography (UPLC) and Ultra High Performance Liquid Chromatography (UHPLC) not only cut time, but also the use of organic solvents, explains Dr. Asif. Additionally, higher selectivity offered by such techniques, in many cases, allows substitution of a solvent such as acetonitrile, which has a higher toxicity with a less toxic solvent such as methanol. “Actually, Ascendia has a program in place to substitute acetonitrile with a more environmentally friendly solvent, methanol, wherever feasible.”
With 50 years of expertise in on-site manufacturing of antibodies and validation/immuno-analysis services, Bethyl Laboratories (a Fortis Life Sciences brand) has been supporting biopharma/biotech industries and academic researchers for their immunoassays-related. In-house quality control (QC) labs validate the antibodies Bethyl makes for several immuno analytical methods, such as western blot, immunoprecipitation, flow cytometry, and immunohistochemistry (IHC). Bethyl’s full-service IHC lab can fix and process tissue for embedding, histological sectioning, and perform immunostaining analysis of a range of analytes. Recently, Bethyl introduced tyramide amplification system-based fluorescent multiplex IHC (mIHC) to its IHC validation/analytical services.
“mIHC is becoming a key tool in understanding cellular interactions in drug discovery and drug development process e.g., immune cells’ interaction with cancer cells in tumor microenvironment,” says Senior Director of Immunohistochemistry and Digital Pathology, Dr Mike Spencer, who oversees the IHC analysis services at Bethyl Laboratories. “As a result, we have invested a lot of effort into expanding our mIHC service, improving efficiency of target panel-building and optimization, and decreasing turnaround times.”
He describes one of the recent services Bethyl provided to a client that needed to stain tissue microarrays containing control and pathological samples to analyze the status of immune infiltrating cells. This was done using a multiplex panel comprised of previously validated antibodies that were translated to the client’s sample type. “We were able to quickly/effectively optimize an antibody panel to determine the T-cell numbers and phenotype several T-cell sub-types within the samples,” he explains. “Our ability to efficiently move from optimization to immunostaining samples with our mIHC expertise saved months of time, precious samples, and other resources for the customer.”
The Bethyl IHC service lab uses automated whole slide scanners for imaging of the stained tissue sections. This allows for analysis of the entire section as opposed to select regions of interest, providing for a more complete picture of markers and their spatial relationships, Dr. Spencer says. This is particularly important, as tissue heterogeneity (e.g., in tumors) is one of the leading concerns in immunostaining analysis. “The automation also allows for consistency and reliability amongst and between samples. With the combination of imaging systems in our lab, we have the capability to image single-stained brightfield images, single-plex fluorescent images, or multiplex images.” These whole slide images are then uploaded to an image database, which the customer uses to view or download their mIHC data for downstream analysis.
Catalent offers a broad array of small and large molecule analytical solutions, from compendial testing to extractables and leachables and cell-based assays. Specialized capabilities include handling of controlled substances, highly-potent compounds, as well as temperature-, light-, pH-, and oxygen-sensitive APIs.
According to Jeff Schwartzenhauer, Analytical Group Leader – Product Development, Catalent, the company has extensive experience and the capabilities to perform analytical testing on a variety of dosage modalities including mRNA, mAbs, cell and gene therapy products, and several dosage forms, including solid oral dosages (capsules, tablets, softgels), inhalation products, and injectables. Catalent also has expertise in characterization, spanning techniques from residue level analysis up to higher order structure (HOS) using state-of-the-art instrumentation across spectroscopy, biophysical, bioassay, and mass spectrometry.
Additionally, Catalent offers in-house identification of visible particulates using modern Fourier transform infrared (FTIR) spectroscopy and Raman microscopes. “Our team of career forensic scientists provides a level of expertise that can rapidly assist in investigations and avoid third-party outsourced testing, which significantly reduces delays,” says Todd Stone, PhD, Director, Analytical Development, Catalent Biologics.
With regard to biologics, Catalent deploys two advanced mass spectrometry (MS) analyses in support of drug substance process development and HOS. “Coverage of host cell protein (HCP) is typically performed using 2D gel assays, but suffers from shortcomings preventing a true measure of the HCP species involved,” says Dr. Stone. “Catalent uses an MS approach, which provides a more thorough assessment, with depth of information on coverage, identity of HCP species, and potential quantitation that outpaces the traditional gel-based method, while shortening the timeline for analysis. For HOS, Catalent uses a covalent label to decorate molecules, followed by high-resolution MS to examine structural modifications because of stresses typically encountered in drug substance or drug product processes.”
As an example, a client with a fast-track biotherapeutic required evidence that a standard HCP kit was sufficient to detect and quantify clearance in its drug substance manufacturing process, describes Dr. Stone. Proceeding with the standard gel-based approach through a third-party lab exceeded their timeline for filing. “However, using the innovative MS-based HCP coverage approach developed and described above, Catalent was able to provide the supporting information to enable the client filing in much less time, and meet the filing deadline,” he says.
Providing information relies on technology. Arvind Ramakrishnan, Director, Lab Automation, Catalent, says technology and analytics offer the ability to access raw data in near-real-time. “Having early access to data could potentially de-risk projects that could go out of specifications, which in turn, saves cost and time,” he says. “Replacing outdated equipment in the lab with smart devices helps partners share data streams with sponsors in near-real-time. The resultant data streams could then be harnessed using platform technologies to not only inform sponsors on status but also positively intervene and salvage runs that are going out of specification. Catalent is actively working on proof-of-concept (PoC) ideas to increase the digital maturity of external projects.”
He adds that automation and analytics approaches using low-code technologies could help generate in silico predictive models, even before running of the actual samples. “Having models to predict outcomes before actual sample runs could help companies avoid failure costs,” Mr. Ramakrishnan says. “Catalent is working with technology partners to help meet this objective through PoC simulation work on “Lab of the Future” platforms.”
When it comes to testing, devices like prefilled syringes, autoinjectors, and pen injectors have specific FDA requirements. For instance, the bulk of the requirements surrounding the physical and mechanical performance of a prefilled syringe to measure attributes such as leakage, break loose, extrusion force, and burst resistance is covered by the ISO 11040 series of standards. Many of these same mechanical and performance measurements for auto and pen injectors are outlined in the ISO 11608 series of standards.
Another key aspect of ISO 11040 for glass Luer syringes is the subject of connectivity. The manufacturing process produces a slippery and slightly irregular Luer taper, which can have issues forming a secure connection with some components. The ISO standard addresses this issue by recommending that certain Luer tests from ISO 80369-7 are conducted to demonstrate adequate connectivity to the same components, which will be attached to the syringe in the actual use situation. ISO 80369-7 replaces ISO 594.
The rapidly growing injectable market has also brought the need for more accurate Container Closure Integrity (CCI) testing to meet USP-NF<1207> deterministic CCI requirements, says Chris Murphy, Marketing Manager, DDL, Inc. The four primary deterministic tests include:
- Helium Mass Spectrometry – best suited for evaluating the inherent integrity of a package system.
- High Voltage Leak Detection – a standard approach for assessing container closure integrity of a nonporous package system.
- Vacuum Decay Testing – applicable to any package containing headspace, including, but not limited to, parenteral vial packages, screw-capped bottles, autoinjectors, and flexible bags or pouches.
- Headspace Analysis – Assessment of package headspace via laser-based analysis techniques provides a quantitative, nondestructive measure of oxygen, carbon dioxide, water vapor, or internal pressure in a nonporous, rigid or non-rigid package’s headspace.
Additional testing, which accounts for drug/device interactions, will likely need to be performed to bring a prefilled syringe product to market includes:
- extractables and leachables;
- USP particulate matter identification and determination;
- ISO 11607 package validation; and
- stability studies to determine product shelf-life.
More and more APIs are entering the clinical phase with no chromophore or possessing a very weak chromophore, says Jerry “Jr.” Mizell, Senior Director of Analytical Services at Metrics Contract Services. “The need for methods that have adequate sensitivity for assay and impurities could pose a substantial issue,” he explains. “Having other means of detection on an HPLC system, such as charged aerosol detection (CAD), refractive index (RI), and mass spectroscopy (MS) will be critical going forward to be successful with challenging compounds.”
As an example, he says Metrics was recently challenged by a clinical project with a very simple API structure containing no chromophore and highly reactive when exposed to water and several other organic solvents. In addition, one of the known impurities and a process intermediate were mutagenic and had to be controlled and analyzed at very low levels (ppm). Several approaches had to be taken for assay and impurity analysis including gas chromatography, RI, and MS. Diluent selection and sample preparation were challenging due to the reactive nature of the active. “But, all challenges were overcome with successful method development and validation for all methods.”
Mr. Mizell adds that MS is also a valuable tool for a CDMO to possess as the FDA’s requirements for nitrosamines is a requirement that all NDA filings must meet. “The same can be said for elemental impurities as it behooves a CDMO to possess ICP-OES and ICP-MS instrumentation,” he says.
“Salubrent Pharma Solutions is a recently formed CDMO focused on supporting the shift toward biologics and personalized medicines, where small batch fill and finish services, combined with continuous batch-fed API processes and just-in-time/direct-to-patient delivery, will ensure patients are never without the life-saving therapies they require,” says Anand Padmanabhan, Director of Analytical Development at Salubrent. “To this end, Salubrent has built its analytical services lab to meet the needs of this changing paradigm.”
In addition to its broader offering of small-molecule analytical testing methods, Salubrent has assembled a team of scientists, and specific instrumentation, focused on large-molecule analytical method development. One specific challenge developing these therapies is the identification of impurities in drug substances and finished goods. Mr. Padmanabhan views gas chromatography (GC) as a vital analytical technique in this effort because of its ability to separate the organic volatile compounds of a sample mixture (typically drug substances) and detect them, thus determining their presence or absence and/or how much is present. “GC is also helpful in the determination of potency, dissolution rate, cleaning verification, etc. in instances where compounds cannot be detected using HPLC UV,” he adds.
Analysis for actives that do not have a UV chromophore cannot be detected using HPLC alone. “In this area, GC is a useful tool, and by adding mass spectrometry (MS), this instrument becomes even more powerful in the identification of impurities,” according to Mr. Padmanabhan. “Overall, a compound is identified via GC-MS not only by comparing its retention time to a standard (GC), but also by using its mass spectrum, making GC in combination with MS an extremely powerful analytical tool.”
GC/MS analysis has endless applications in material testing, identification, and certification. Identification of organic volatile impurities is one of the main applications. In the pharmaceutical industry, GC/MS is used in research and development, production, and quality control. In medicinal chemistry, GC/MS is used in the synthesis and characterization of compounds and in pharmaceutical biotechnology.
“Salubrent has the GCHS and GC/MS instruments analysis capabilities and expertise necessary to handle these types of complex projects,” says Mr. Padmanabhan, adding that he views Salubrent’s ELISA testing, Malvern Mastersizer particle size analysis, and HPLC with PDA impurity and assay potency quantitation capabilities as key to supporting the industry’s increasing focus on biologics and personalized therapies.
SGS has three centers of excellence in North America that offer analytical testing for biologics – located in Pennsylvania, Illinois, and Ontario, Canada – providing services at every stage of a product’s lifecycle, from early-phase cell bank safety assessment and product characterization to later phase method development and final phase GMP product release.
For the last seven years, SGS’s chemistry laboratory in Lincolnshire, IL, has extensively used Pinnacle PCX, a post-column derivatization system from Pickering Laboratories that performs analysis of amino acids for individual raw materials as well as the evaluation of small peptides. This instrument replaced the thin layer chromatography (TLC) test used in the past to monitor ninhydrin positive substances, explains Natalia Belikova, PhD, Analytical Services Director, SGS. “High Performance Liquid Chromatography instrument (HPLC) technology is more specific than TLC, has better sensitivity, is faster, and costs less. This instrumentation was also successfully used for identification tests for small peptides, when identification is based on molar ratios of known amino acids and presence/absence of other known amino acids.”
Additionally, the SGS laboratory has an X-ray powder diffractogram D2-phaser (XRPD) from Bruker that is used extensively for the identification of different known polymorphic forms of small molecules. “It also allows us to evaluate API purity,” she says. “Several clients have asked us to run confirmatory testing that polymorphic structure of active pharmaceuticals ingredients does not change when an API is incorporated into the final drug product during the manufacturing process. This methodology is useful when clients ask us to evaluate if stability storage (under International Conference on Harmonization or accelerated studies) affects the polymorphic form of active ingredient.”
Triclinic Labs, Inc.: Chemical Analysis & Solid-State Development
Triclinic Labs provides both cGMP and non-cGMP materials characterization of organic and inorganic molecules. The lab has a DEA-Controlled Substance Registration for Schedule I-V compounds and offers a variety of techniques, including PXRD, NMR, TGA, DSC, DVS, SEM, EDX, IR, FT-IR, Raman (Dispersive, Low Frequency), optical, digital, hot-stage, and polarized microscopy, optical rotation, contact angle, particle size and quantity analysis, HPLC, GC with headspace, ICP-MS, and a variety of other techniques for identifying chemical composition, contaminants, and unknown substances.
Determination of crystalline in amorphous mixtures is a requirement of the FDA to demonstrate control in chemical manufacturing (CMC). Aeri Park, PhD, Chief Scientific Officer, Triclinic Labs, explains that Triclinic has developed innumerable quantitative methods to determine the presence or absence of crystalline polymorphs in solid mixtures at extremely low levels of detection, and well below the FDA’s typical 5% w/w request. These methods are then used to release materials for pharmaceutical drug product manufacturing.
- United States $5.55 Billion Pharmaceutical Analytical Testing Outsourcing Market – Size, Share & Trends Forecast to 2027, Research and Markets, Feb. 1, 2021, https://www.globenewswire.com/en/news-release/2021/02/01/2167066/28124/en/United-States-5-55-Billion-Pharmaceutical-Analytical-Testing-Outsourcing-Market-Size-Share-Trends-Forecast-to-2027.html.
- Global Pharmaceutical Analytical Testing Outsourcing Market Analysis Report 2021-2028, ResearchAndMarkets.com, Oct. 12, 2021, https://www.businesswire.com/ news/home/20211012006008/en/Global-Pharmaceutical-Analytical-Testing-Outsourcing-Market-Analysis-Report-2021-2028.
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