Chromatography – A Journey Through Time
Follow the evolution of process chromatography from the proverbial dark ages to the latest innovations.
This article is posted on our Science Snippets Blog
Robust purification procedures are essential to the efficient production of safe biotherapies. Most biopharmaceutical manufacturers employ at least one chromatography step within their downstream bioprocess. Moreover, the industry relies on various methods of chromatography, classified by features like chemistry, matrix, and mode. Yet, have you ever wondered how the science evolved to yield the diverse toolbox we have today?
Join us as we explore the – often surprising - history of chromatography.
But First – Let’s Review Some Terms and Definitions
Chromatography describes any technique used to separate components of a mixture by passing them through a stationary phase (such as a membrane, resin, or monolithic matrix).
Separation occurs because the components of the sample interact differently with the stationary phase, according to features like size, charge, hydrophobicity, and affinity for a specific ligand.
The goal is to isolate sufficient quantities of a target component (preparative chromatography) and | or quantify and analyze the features of the individual components (analytical chromatography).
Discover more terms and phrases in our Chromatography Glossary
The Dark Ages
The invention of chromatography is typically traced back to the early 1900s. However, scientific advances tend to emerge gradually, with iterative improvements to existing technologies and knowledge. This makes defining an exact timepoint quite tricky.
Some claim that the earliest hints of chromatography can be traced back significantly further than the 1900s. In the book “Chapters in the Evolution of Chromatography,” analytical chemist Leslie S. Ettre asks, “Was Moses The First Chromatographer?” This strange idea comes from the story in the Book of Exodus, in which Moses throws a log into a bitter water source, sweetening it enough to make it drinkable. This phenomenon represents an (albeit very simple) illustration of the principles of chromatographic adsorption!
The earliest hints of applied chromatography principles emerged in the 1800s. One example is the work of German Chemist Friedlieb Ferdinand Runge, who had a keen interest in the behavior of colored dyes. He began to note patterns when they were blotted on filter paper, which he used to monitor chemical reactions. Although there is controversy surrounding Runge’s role in the evolution of chromatography, it is clear that some features of his work could certainly be interpreted as a predecessor to paper chromatography.
The Golden Age
The invention of what we now know as chromatography is typically attributed to Russian botanist Mikhail Tsvet. He developed the technique in the early 1900s and used it to separate plant pigments. It wasn’t until the 1930s and 1940s that chromatography methods were further developed, making the technology more widely applicable.
Archer Martin and Richard Laurence Millington Synge later won the Nobel Prize in Chemistry for the invention of partition chromatography. This recognition put the spotlight on the technique and set the stage for significant progress, including the broader adoption of gas chromatography and high-performance liquid chromatography (HPLC). HPLC was further refined and employed in various applications throughout the 1960s.
The primary targets at this time were small molecules and chemically well-defined mixtures. The application to more complex biological systems was evolving more slowly. The gentle nature of conditions in aqueous chromatography eventually supported the application of chromatography for the separation of biologics.
The Renaissance
The first commercial HPLC machine was produced in 1967. During the next twenty years, the HPLC toolbox was developed to facilitate the separation of increasingly similar compounds. Supported by the commercial availability of carbohydrate-based chromatography matrices and improved detection | analysis modules, the 1960s also sparked a revolution in protein chromatography. These advances enabled the careful separation of more diverse and complex molecules through technologies such as reverse-phase, hydrophobic interaction, and affinity chromatography.
By the 1980s, HPLC was commonly used in laboratories worldwide. Iterative improvements in sensitivity, speed, accessibility, and resolution continue to be made.
The Industrial Revolution
Armed with a diverse choice of chromatography approaches, scientists encountered a new hurdle: how can we scale up separation procedures to serve the biopharmaceutical industry?
Chromatography is an invaluable step in isolating meaningful yields of biotherapies, such as monoclonal antibodies (mAbs). However, simply increasing the size | volume of a chromatography device leads to issues with matrix stability, pressure limits, and column structures.
New column features and enhanced binding capacity are helping biomanufacturers increase their throughput while retaining the speed and resolution of their separation processes. Scale-up is also supported by integrated technologies that enable the integration of both single-use technologies and process automation, increasing consistency and throughput of the entire downstream process.
The Information Age
Most recent advances in chromatography have been driven by the needs of the sectors it serves, including the biopharmaceutical industry. Here, there is a significant drive to reduce the time-to-clinic | time-to-market, produce higher yields, keep up with new modalities, and limit facility footprints. Chromatography methods must be sensitive, fast, and efficient to keep up with these demands,
Technologies such as multi-column chromatography (MCC) are gaining traction in the bioprocessing industry. These continuous approaches enable continuous and intensified processing, allowing purification to be carried out more efficiently and reducing costs. Analytic sensors with increased sensitivity and more comprehensive, user-friendly analysis platforms deliver new process | product insights. These developments promote improved quality standards and consistency between batches.
The Future of Process Chromatography
The innovations of the last century have shaped the field of modern-day chromatography. Used in laboratories and bioprocesses worldwide, chromatography techniques have been pivotal in establishing countless biologics and therapies that are fundamental to patient care.
The next era of chromatography technology will lie in the purification of novel modalities like cell and gene therapies, bringing new drugs and treatments to market at an unprecedented rate.
References
(1) Ettre, L. S., & Hinshaw, J. V. (2008). Chapters in the evolution of chromatography. In Chapters in the Evolution of Chromatography. https://doi.org/10.1142/P529
(2) Celia Henry Arnaud. (2016). 50 years of HPLC. C&EN Global Enterprise, 94(24), 28–33. https://doi.org/10.1021/cen-09424-cover1
(3) Majors, R. E. (2015). Historical developments in HPLC and UHPLC column technology: The past 25 years. LC-GC North America, 33(11), 818–840. https://www.chromatographyonline.com/view/historical-developments-hplc-and-uhplc-column-technology-past-25-years
(4) Curling, J., & Gottschalk, U. (2007). Process chromatography: Five decades of innovation. In BioPharm International (Vol. 20, Issue 10, pp. 70–94). https://www.biopharminternational.com/view/history-chromatography-process-chromatography-five-decades-innovation