Miniaturised Purification with Antha and Tecan Te-Chrom™: Uniting Robust Execution With Flexible Planning and Comprehensive Data Analysis

Purification of target compounds is a key step in the production of final, market-ready products in bioprocessing. Various industries, from pharma and cell therapies to food and flavourings, rely on column chromatography in its many modalities and volumes, ranging from miniaturised columns of less than 1 mL used for process development to columns exceeding tens of litres user for manufacturing.

To achieve high-throughput purification, our partner Tecan developed a miniaturised column purification system called Te-Chrom™. It can be installed on a Tecan Freedom EVO® or Tecan Fluent® liquid handling platforms and uses resins with volumes between 50 and 600 µL packed in miniaturised columns, like the Repligen OPUS® Robocolumns®.

Taking advantage of the precision and accuracy of automated liquid handling to handle fractions of less than 50 µL, this system allows for multiplexed purification while saving precious sample volume. Such systems typically also feature an on-deck plate reader for taking absorbance measurements of flow-through liquids to determine sample concentration.

Leveraging the Full Potential of Miniaturised Purification with Antha

In conjunction with miniaturised bioreactors, these miniaturised purification systems using RoboColumns® form the cornerstone of High Throughput Process Development (HTPD). While this technique has been around for more than a decade and has already unlocked new opportunities in bioprocessing, there are still limitations in the planning and execution of sophisticated experiments where, for example, a range of buffers are being used in the same chromatography stage.

It can often be cumbersome and difficult to handle and track a large number of samples and buffers, set up sophisticated optimisations of purification conditions (sometimes involving Design of Experiments (DoE)), and structure experimental data for further analysis. In particular, data augmentation, normalisation, and background subtraction are often manual, inflexible, and error-prone.

With these problems in mind, Synthace has recently developed a protocol to facilitate the planning, verification, execution, and data analysis of such workflows. The protocol is executed using our software platform Antha, which has been designed to empower scientists to use lab automation to its full potential.

In a recent online demo, our Scientific Development Consultant, Dr Milena Stanković, demonstrated this new miniaturised purification protocol in Antha, designed to complement and expand on the current Tecan Te-Chrom™ offering.

Watch the trailer ↓

Antha’s End-to-End Process for Miniaturised Purification: from Experimental Design to Data Analysis

Antha is a cloud-based platform that automatically translates standard experimental instructions given by the user (i.e. inputs in everyday language) into a set of liquid handling instructions for the automation equipment, prompting it to execute the experiment and thereby eliminating the requirement for user programming knowledge.

Experimental inputs (sample names, concentrations and extinction coefficients, resin and buffer metadata, purification conditions like column volumes and residence times) are defined using Antha’s intuitive interface.

The experimental design is then automatically tested for feasibility by Antha’s simulation engine. As part of this in silico test, Antha provides a detailed overview of each liquid handling step, allowing scientists to verify their experiments prior to execution and iterate over the design if needed.

All this can be done from the comfort of your desk, in the office or at home, without the need to occupy the robot to plan the workflow.

Furthermore, within Antha’s user-friendly interface, scientists can select absorbance measurement protocols on the associated plate reader, such as the Tecan Infinite® M200 Pro. Because Antha has calculated all necessary liquid handling steps, it can augment the plate reader data and indicate which absorbance measurement corresponds to which chromatography phase automatically.

After the workflow is executed and the data is collected, Antha then generates visualisations (including pseudochromatograms) for analysis of the collected data by performing calculations of the flow-through volume, sample concentration, and other parameters such as mass and yield.

How Can You and Your Team Benefit from Miniaturised Purification with Antha and Te-Chrom™?

Antha lifts the massive burden facing biologists today, where data collection, alignment, and normalisation are often tedious and error-prone. Even if in-house software solutions exist, these are often inflexible, difficult to maintain, and require advanced programming skills.

Antha integrates with automation equipment to allow scientists with no programming background to perform end-to-end experiments, such as the newly available miniaturised purification workflows. Users are given highly granular control over the process and can now define their experimental conditions with ease.

Another important advantage of using Antha for purification experiments on the Tecan Te-Chrom™ system is that the Antha platform’s general liquid handling functionality can be used to prepare samples for further downstream analytics, such as ELISA, electrophoresis, or HPLC. Within Antha’s analysis page, relevant samples can be automatically pooled or cherry-picked based on, for example, a concentration measurement.

Design of Experiment (DoE) files from JMP or Design Expert are also inherently supported in the purification protocol. Antha’s in silico testing and liquid handling preview prior to execution allows users to trace their samples throughout the workflow and empowers them to embrace sophisticated optimisations with confidence.

Furthermore, because Antha keeps track of the log files, the liquid classes, and labware definition at run-time, experiments become fully traceable: Antha keeps all its history in its own searchable archive. In addition, Antha’s comprehensive data analysis prevents errors and relieves scientists from manual intervention, thus saving precious time and resources.

To see Antha in action and learn how it can empower your high-throughput miniaturised purification assays on the Tecan Te-Chrom™ system, watch the full Antha demo.

Watch the full demo here