The Evolution of Biopharma Analysis

The Evolution of BIOPHARMA ANALYSIS What are BIOPHARMACEUTICALS? Biopharmaceuticals – also known as biotherapeutics – are medications derived from biological sources via biotechnological processes. Sources can include: Genetically modified Plants Microorganisms cells or organisms 1980 The term “biopharmaceuticals” 1980 was first coined. 1982 The first biopharmaceutical, recombinant human insulin, was approved for human use by the US Food and Drug Administration (FDA). 2024 The biopharmaceutical market is now worth an estimated $1.5 trillion. Current therapeutic uses of BIOPHARMACEUTICALS Vaccines Cell therapies Gene therapies Tissues Monoclonal Whole blood antibodies (mAbs) components Antigens Hormones Xenotransplantation Enzymes Immunosera Cytokines products BIOPHARMACEUTICALS vs small-molecule drugs Biopharmaceuticals are very different to small-molecule (i.e., synthetic) drugs. Synthetic Biopharmaceutical Origin Synthetic Living organisms Small molecules of low Large, complex molecules Structure molecular weight with well with high molecular weight defined structures Chemical synthesis Manufacturing Biotechnological workflows workflows Biopharmaceuticals carry several advantages over synthetic drugs: they target specific molecules, reduce the risk of side effects and have high specificity and activity. They are also much more complex and have to meet rigorous requirements for quality and safety. The composition, quality, stability and safety of complex biotherapeutics must be assessed throughout the development pipeline, which requires a wide range of analytical techniques. Milestones in BIOTHERAPEUTIC ANALYSIS Let’s explore some of the many technological and regulatory milestones that have shaped modern biopharmaceutical analysis: Early 1900s THE BEGINNING OF BIOLOGICS REGULATION The US Congress passed the 1902 Biologics Control Act, which gave the government control over the processes used to produce biological products. Under this Act, the Hygienic Laboratory of the Public Health and Marine Hospital Service issued regulations and oversaw laboratory inspections in manufacturing facilities. The Hygienic Laboratory was renamed the National Institutes of Health (NIH) in 1948. Biologics control remained part of the NIH until it was transferred to the FDA in 1972. 1950s - 1960s EARLY CHROMATOGRAPHY TECHNIQUES EMERGE Chromatography is a major tool used in downstream processing of biotherapeutics. It can be used alone or coupled with mass spectrometry (MS) and other techniques. The invention of chromatography is attributed to Russian scientist Mikhail Tsvet in the early 1900s. But during the 1950s and 60s, several milestone innovations occurred that paved the way for the suite of chromatography tools now used for biopharmaceutical separation and purification. Such tools include, but are not limited to: Affinity chromatography (AC) The term “affinity chromatography” was used in the literature for the first time in 1968. Exploits interactions between a protein and a AC was a critical discovery for the ligand attached to a resin. emerging biopharma industry – over 50 years later – it is one of the Ligand most commonly used methods for antibody purification. Protein of interest Hydrophobic interaction chromatography Size exclusion / gel filtration chromatography Separates proteins based on hydrophobicity. Separates proteins based on size by passing them through a column filled with porous beads. Porous Large Small beads proteins proteins Hydrophobic Protein of ligand interest Ion chromatography as we know it Ion exchange today took many years to develop. chromatography (IEC) In the 1950s and 60s, researchers refined the technique to improve the separation and purification of Separates proteins based biomolecules such as proteins, on their charge. peptides and nucleic acids. Anionic protein of interest IEC is a key tool in modern biopharma analysis for the Cathionic stationary phase characterization of charge variants of therapeutic proteins. SCIENTIST START TO USE SPECTROSCOPY TO ANALYZE COMPLEX BIOMOLECULES In the 1950s, scientists started to apply nuclear magnetic resonance (NMR) spectroscopy to study small biomolecules like amino acids and ribonucleases. Over the following decades, NMR spectroscopy was used routinely in small molecule pharmaceutical development for the assessment of chemical structure, purity and stability, but less so in biopharmaceutical development. Recently, however, benchtop NMR spectrometry is being used in biopharma to indirectly analyze the properties of therapeutic proteins, for example protein concentration or aggregation. 1970s FURTHER INNOVATIONS IN CHROMATOGRAPHY The introduction of ligands such as coenzymes, lectins, nucleic acids, metal chelates, triazine dyes, Protein A and heparin, further enhanced AC. Protein A AC became a cornerstone technique in therapeutic antibody production, and is also widely used for its effective clearance of impurities, high- and low-molecular weight species, host cell proteins (HCPs) and DNA. Many other advancements in chromatography have occurred since the 1970s that benefit biopharma analysis, such as the increasing sophistication of high-performance liquid chromatography, enabling faster and increasingly efficient separation of a broader range compounds with higher resolution, accuracy and reproducibility. THE 1975 ASIMOLAR CONFERENCE PROMPTS A RISE IN RECOMBINANT DNA RESEARCH One of the first organized discussions about the safety and ethical implications of genetic engineering, particularly recombinant DNA (rDNA) technology, which led to the first guidelines on its research and use. Many consider the Asimolar conference to be a key moment in history for the advent of biopharmaceuticals and their regulation. INVENTION OF THE ENZYME LINKED IMMUNOSORBENT ASSAY (ELISA) TECHNIQUE In 1971, Eva Engvall and Peter Perlman first described the ELISA test. It remains the gold standard method for detecting HCPs, which can cause unwanted immunogenic responses, in biopharmaceutical impurity screening. 1980 and 1990s MS EMERGES AS A TOOL TO STUDY PROTEINS, WITH IMPLICATIONS FOR BIOPHARMA MS, a type of chemical analysis that measures the mass-to-charge ratio(m/z) of atoms and/or molecules in a sample, had been invented, commercialized and coupled with other techniques such as gas chromatography prior to the 1980s. The introduction of electrospray ionization and matrix-assisted laser desorption/ionization during this decade significantly expanded the types of analytes that could be subjected to MS analysis – including proteins. One of the earliest used of MS in biopharma analysis was to confirm the amino acid sequence of recombinant proteins. Advances in MS technology over the decades led it to become an indispensable tool in biopharma analysis, particularly when coupled with liquid chromatography (LC). Various different types of LC MS can now be used for quantification, quality analysis, impurity profiling, structure–function analysis, bioanalysis and clinical analysis of biotherapeutics, among other applications. FDA APPROVES FIRST RECOMBINANTLY-PRODUCED BIOTHERAPEUTIC The FDA’s approval of recombinant human insulin in 1982 marked the acceptance of recombinantly produced biotherapeutics, heralding a “new era” in the biopharma industry and the application and modification of existing analytical tools to explore the quality, safety and efficacy of these new drugs. The FDA Center for Drugs and Biologics was also established in 1982 to streamline and improve regulatory oversight for both pharmaceuticals and biologics. It was later separated into two centers: the Center for Drug Evaluation and Research and the Center for Biologics Evaluation and Research, which continue to operate as distinct entities within the FDA. INVENTION OF THE POLYMERASE CHAIN REACTION (PCR) TECHNIQUE In 1984 Kary Mullis invented the PCR technique, enabling the amplification of DNA, which became fundamental to biological science. Later adaptations of PCR, including the development of real-time PCR and digital PCR, enhanced biopharmaceutical analysis by creating new ways to interrogate bioprocesses and the product itself to ensure high quality. Monitoring the genetic stability of production cell lines Absolute quantification of Residual host cell DNA viral vectors analysis PCR-based Detecting microbial Monitoring antibody methods can be contamination stability used for: ADVANCES IN SPECTROSCOPY Some of the earliest applications of spectroscopy in bioprocessing occurred in the 1980s, such as the first reported use of Raman spectroscopy. Raman spectroscopy would become recognized as a process analytical technology (PAT) tool in biopharmaceutical processing in the early 2000s and is used to monitor cell cultures, ensure raw material quality and optimize biomanufacturing workflows. In the 1980s, researchers also started to use Fourier transform infrared (FTIR) spectroscopy to assess the secondary structure of therapeutic proteins, such as insulin, enhancing our understanding of protein stability. THE INTRODUCTION OF CAPILLARY ELECTROPHORESIS Though the origins of electrophoresis trace back to the early 1900s, modern capillary electrophoresis (CE) was introduced by Jorgenson and Lukacs in 1981, leading to its popularization. Since the 1980s, several CE-based methods have been developed, such as CE sodium dodecyl sulphate , capillary zone electrophoresis and imaged capillary isoelectric focusing. CE-based methods have become a staple tool for the characterization and QC of a wide variety biomolecules, such as mAbs, fusion proteins and vaccines, due to their versatility, low sample consumption, accuracy and potential for automation. CE separation can also be coupled to other detection techniques for a variety of biopharmaceutical analysis applications, such as laser-induced fluorescence for capsid protein purity analysis in gene therapy products, or MS for the characterization of therapeutic proteins. 2000s onwards INTRODUCTION OF NGS The “genomic revolution” began in the early 2000s, paving the way for novel biotherapeutics based on genetic engineering and editing. Next-generation sequencing (NGS) tools became faster and more affordable. NGS can be used to monitor environmental contaminants in biopharma manufacturing processes, demonstrate genetic stability in cell lines and confirm the integrity of gene therapy products. The International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) recently published new guidelines that highlight the benefits of NGS assays vs in vivo assays for detecting viral contamination in products produced from in vitro cell culture using rDNA technologies. US FDA LAUNCHED CURRENT GOOD MANUFACTURING PRACTICE (cGMP) IN 2002 The FDA’s launched the cGMP guidelines – a framework for the biotherapeutic to ensure products are manufactured according to quality standards and in accordance with their intended use. FDA PUBLISHED PAT FRAMEWORK GUIDANCE IN 2004 PAT is a framework created to ensure quality by integrating real-time monitoring and control of critical process parameters during production. PAT solutions introduce a feedback loop to the manufacturing process, which supports higher efficiency, consistent product quality and faster time-to-market. Different ways to implement PAT solutions in bioprocess monitoring: On-line Off-line A C Analyzer In-line At-line B D Analyzer THE RISE OF AI AND AUTOMATION Artificial intelligence and automation could transform biopharma manufacturing, streamlining processes from design to quality control, reducing labor costs, enhancing production efficiency, ensuring regulatory compliance and reducing waste. Their increased adoption is poised to drive more sustainable and optimized operations. Sponsored by