Chromatography Counselling - Let’s Get Analytical
What is it? How is it different from preparative chromatography? When should you use each method, and should you really be using both? Here, we answer all your burning questions on analytical chromatography.
This article is posted on our Science Snippets Blog
Preparative and analytical chromatography of large biomolecules can be performed on a monolithic matrix
There are various types of chromatography, typically classified by chemistry, the structure of the stationary phase, the physical state of the mobile phase, methodology, and device. But first, we can categorize approaches as either “preparative” or “analytical,” depending on their intended purpose. That is, what are you trying to achieve?
Preparative Chromatography vs. Analytical Chromatography
In biomanufacturing, preparative chromatography aims to isolate and purify sufficient yields of a target component - such as a protein, nucleic acid, or viral particle - in a mixture. The purified target is used in downstream applications such as the creation of biotherapeutics.
On the other hand, analytical chromatography gathers qualitative and quantitative information. In the biopharmaceutical industry, analytical chromatography is used to separate and quantify the target molecule from other isoforms, aggregates, and contaminants. This allows biomanufacturers to better understand their target molecule in a complex mixture, enabling process control and optimization. It also ensures quality control and product safety by monitoring and providing insights into the precise components of the sample at various process stages.
The general principles of these methods are the same. Still, there are critical differences that allow each type to serve its purpose. These are outlined in Figure 1.
Figure 1 – Analytical and preparative chromatography rely on the same principles, but have distinct aims and features.
CIMac™ Analytical Columns possess all the characteristics of CIM® monoliths at analytical scale.
Both types of chromatography rely on the same principles, including a range of separation chemistries and methodologies. The fundamental difference between analytical and preparative chromatography is the end goal. To achieve its goal, each has distinct features and requirements that affect its performance.
Let’s take a closer look at how some operational differences ensure the effective performance of each strategy in biomanufacturing.
Scale
Typically, analytical chromatography occurs on a small scale. The goal is to separate components and collect a small amount of material, just enough to perform a detailed analysis and determine relative quantities of the relevant components. In contrast, preparative chromatography is carried out on a larger scale. Typically, the eventual goal is to commercially produce a biomolecule, so the system must be able to handle production-scale purification processes.
The PATfix® HPLC system, powered by PATfix® software, represents a user-friendly toolbox for automated analysis of chromatographic data sets.
As you might expect, the sample volumes for preparative chromatography are significantly higher than those required for analytical chromatography (typically in the range of milliliters to liters, as opposed to microliters, respectively).
Column Features
Related to scale and application, column diameter, length, and matrix particle sizes are also different between the two approaches. Column dimensions are smaller for analytical chromatography compared to preparative chromatography. The particle size in the chromatography matrix is smaller in analytical columns, usually five microns or less, compared to ten microns and above in preparative columns. The combination of smaller column dimensions and smaller particle sizes reduces the flow rates, creating sharper, more defined peaks necessary for proper analytics. Larger particles facilitate higher flow rates, leading to broadened peaks, less of an issue in preparative chromatography.
Technologies
While not so critical in preparative chromatography, sensitive sensors, detectors, and analysis platforms are essential components of an analytical process. These tools are implemented to facilitate robust data collection and analysis, supporting proper process control and addressing quality needs.
At-line analytical systems, such as PATfix®, is critical for successful analytical chromatography. Supporting software is essential for the proper monitoring, recording, and evaluation of separation processes and final products. Immediately available and accurate information supports in-process control and process development, reducing the risk of failure and speeding up product release.
The Right Chromatography Tools and the Right Skills for the Right Tasks
Sartorius has a variety of tools to meet your analytical chromatography needs, allowing you to delve deeper into your downstream purification process and achieve rapid product release.
Our PATfix® HPLC system, in combination with our CIMac™ Analytical Columns, allows you to monitor and control critical process parameters (CPPs) and critical quality attributes (CQAs) of your product. Our tools are particularly suitable for the next generation of modalities, including large biomolecules like mRNA therapies and viral vectors.
Explore our chromatography portfolio and find out how you can optimize your analytical chromatography process.
