Protocol Details: Miniaturized Purification with Synthace and Te‑Chrom™

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Why this protocol?

Synthace for miniaturized purification provides an end-to-end solution for planning, executing and structuring data for high throughput purification workflows to accelerate your process development.

Batch processing users will benefit from Synthace's data structuring and traceability of runs.

Process optimization scientists will appreciate Synthace's flexibility, enabling sophisticated Design of Experiments.

Pain points and current processes

  • Current practices

    With Synthace purification

  • Experiment Inputs

    Current practices

    • Can’t easily link metadata to samples, buffers and additives with Excel or Te-Chrom™ software
    • Can’t define more complex experimental designs with simple imports

    With Synthace purification

    • Easily link metadata to samples, buffers and additives with Synthace
    • Undertake more complex experimental designs, such as DOE
  • Experiment Planning

    Current practices

    • Planning must be done on attached PC, and can’t be done when the robot is running
    • Difficult to change experimental parameters in the EVOware® UI
    • Excel spreadsheets with sophisticated macros are inflexible and difficult for wider team to use

    With Synthace purification

    • Plan your experiment remotely, from anywhere
    • Flexibly change experimental parameter with Synthace’s intuitive interface
    • Share flexible workflows with team members
  • Experiment Execution

    Current practices

    • User needs to understand liquid handling hardware details to modify execution
    • Excel spreadsheets only create worklists and can’t integrate analytical device data

    With Synthace purification

    • Simply upload the instruction file to SynthaceHub and Synthace orchestrates the hardware
    • Synthace handles the execution of the experiment, providing more than just a worklist
  • Data Aggregation

    Current practices

    • No simple way to align plate reader data with sample metadata and chromatography phases

    With Synthace purification

    • Synthace enables full sample provenance. Plate reader data, sample metadata and chromatography phases are all tracked
  • Data Normalization

    Current practices

    • No simple way to generate chromatograms
    • Extra software package in the loop if using plate reader software to normalize data

    With Synthace purification

    • Synthace creates chromatograms
    • No extra analysis software needed

Protocol walkthrough

Basic concepts

Translate human readable actions into machine instructions

Simple building blocks...

Elements are a key concept of the Synthace platform. They translate human readable actions into machine instructions. You can define how the experiment will be performed through the element input parameters.

Elements form the visual user interface to Synthace's planning engine. Synthace interprets your requirements and translates these into complex robot instructions. Input volumes are also calculated and the labware needed is determined.

...translated into complex robot instructions

You can create workflows in Synthace by connecting elements together to specify the flow of physical materials and information.

A simple workflow example is connecting an element where liquids and their properties are defined (e.g liquid names, concentrations, pH, and so on) with an element that aliquots these liquids onto a plate as desired.

Define experiment

Easily define and modify your workflows

Define RoboColumn® Plate — where you specify resin types and volumes.

Define Chromatography Step — where you define the name of the step (e.g. Wash, Load, Elute), the inlet buffer, flow rate, volumes to load, and whether fractions need to be collected.

This is an example of a standard purification workflow including an absorbance measurement step, as viewed in Synthace’s Workflow Builder. We start by defining the RoboColumns®, followed by several repeating elements to define chromatography steps as well as buffers.

Synthace also allows you to easily create SOPs using the template feature. With templates, you can easily modify a few parameters of interest — such as number of samples to process, for example — while the majority of the workflow remains unaltered. This ensures reproducibility and maximizes ease of use.

Once a workflow is defined, you can simulate it from the comfort of your desk. The simulation invokes sophisticated algorithms that calculate all liquid handling steps and also the required labware and source liquids.

After a handful of seconds, you will be able to see the result of your simulated workflow. Should you not be satisfied with the resulting plan, you can easily go back, edit your workflow and iterate.

Preview in silico

Preview your runs in detail at your desk

Synthace's Execution Overview presents the required labware as well as a shopping list of your reagents. You can also see the devices on which you are going to run your workflow.

Synthace’s Execution Preview shows a layout of the Tecan Freedom EVO® liquid handler deck, including the Te-Chrom™ system, and clearly shows where labware needs to be placed.

This Preview also shows every liquid handling action in order of execution with a useful step-by-step time estimate. This minimizes the chances of errors, and gives you an immediate estimate of walk-away time. Hovering over individual transfers will indicate the selected liquid handling policies or liquid classes.

Automated execution

Easily send jobs to your lab for execution

Once you’re happy with your experiment planning, you can schedule it for execution. This will send your set of instructions from the cloud to your Freedom EVO®.

The recipient of these instructions is an app called SynthaceHub, installed on the computer connected to your Freedom EVO®. SynthaceHub uses the Freedom EVOware® API to translate the instructions generated by Synthace into Freedom EVOware® instructions.

Output

Data structuring and export is automated with Synthace

Once the experiment has completed, your data is securely sent back to the Synthace Cloud. Plate reader data is aligned with the sample liquid handling steps, and normalized for pathlength and blank corrected where appropriate. 

You can conveniently download your data from the Results page for analysis in your favourite software.

You can also view the log and config files associated with your run for full traceability. Barcodes are also displayed if read.

By opening the Data Visualization tools, you will be able to interact with your data.

The Chromatogram tab of Synthace's Data Visualization tools displays data from a standard purification. 

Different columns can be easily selected and the various chromatography steps are indicated on the absorbance graph. Absorbance spectra for all acquired wavelengths are displayed

The save button allows you to download all the plots you see on screen as image files for reporting purposes.

The Analysis tab gives you a granular view of the structured data from the collection plates containing each fraction.

Using the range of views and filters available, you can normalize and inspect your data set without any need for manual manipulation in Excel.

The Analysis setup section allows you to define which data you want to use to correct for pathlength and background absorbance.

The Plate layout section gives you an interactive, graphical view of absorbance, corrected values, pathlength, volume, concentration and quantity, where you can also filter by chromatography stage.

Finally, the Yield data plot at the bottom of the page gives you an instant view of the aggregated data from the elute fractions from each column.

Cherry picking

Accurately select wells for downstream uses

Minimize risk of human error

If you’re looking for an easy method for fraction pooling, the range selector in the Analysis tab of Synthace’s Data Visualization tools allows you to quickly and accurately select wells for cherry picking, for example by specifying the measured concentration range.

This view minimizes risk of human error by ensuring that you’re selecting wells of your desired concentration range.

You simply have to click on the “Cherry-pick selected wells in Synthace” link to proceed.

Select wells automatically or manually

The wells you have selected in the Analysis tab are automatically shown in Synthace’s Cherry Picker tool. Wells can also be manually selected whenever you need to reformat a plate.

Furthermore, you can simply wire any element from Synthace’s standard element set onto the output of the Cherry Picker, for example to further dilute your samples.

ELN interface

Use your existing inventory systems

Interfaces with a number of electronic lab notebooks (ELN) are available if you’d like to use your existing inventory systems to select resins, buffers and samples for your purification workflows.

Here we show an example interface for input selection in the IDBS E-WorkBook.

Once you have the inputs selected and exported as a CSV file from your ELN, you can easily upload those inputs directly into your purification workflow in the Synthace Workflow Editor.

Track your experiments

Clear and searchable status overview

Search, collaborate and share

Synthace conveniently presents a clear and searchable overview of the status of all your experiments, whether they are simulated, scheduled or executed. 

You may also view your co-workers’ experiments to enhance collaboration between teams. Workflows can also be easily shared within your organization, increasing consistency and reproducibility.

As with all modern cloud-based software, nobody outside your organization will be able to access any of your data.

Simulations list in Antha

FAQs

Synthace for Miniaturized Purification is recommended for use with Tecan liquid handlers with Te-Chrom™/Te-Chrom™ Shuttle and RoboColumns® from major vendors.

Synthace provides a wide range of integration options for analytical instruments. We have direct integration with plate readers and indirect integration with downstream analytics such as qPCR and ELISA. If you’d like to enquire about specific processes for your lab, please drop us an email or use the “Request a demo” button below.

No, Synthace supports all resins, as well as both long and short RoboColumns®.

Yes, DOE is core to our application. Synthace currently supports processing DOE design files made in JMP®, Design-Expert®, or any CSV-like file.

The Synthace planning engine parses the high-level inputs specified in your workflow, including the experimental design, selected labware and deck layout. The planning engine converts these instructions into low-level instructions such as the liquid handling steps and the exact labware positions on your predefined deck layout.

Yes, custom deck layouts can be chosen in Synthace. The user can define multiple deck configurations for the same liquid handler model, such as the Tecan Freedom EVO®.

Synthace will work out the changes necessary to implement your workflow. You simply need to upload your deck configuration and re-simulate — Synthace will use your new worktable layout to determine the necessary instructions.

Yes, you can — this can include activating a pump, should you have one.

Our Field Application Scientists will learn your processes to set up your initial workflows during deployment, but phases can easily be added, removed, and reconfigured and new workflows can also be built using the workflow buider.

You can select the liquid handling policy for each of the chromatography stages individually. Different samples can have different policies, and these policies are linked to liquid classes on your Tecan Freedom EVO®.

Synthace generates Freedom EVOware® scripts that the user can download and inspect.

Troubleshooting is mostly done in Freedom EVOware® and we currently do not automate responses to errors generated during a run. Instead, our working principle is to prevent errors pre-run. Synthace’s planning and simulation capabilities work to prevent a range of errors before executing in Freedom EVOware®, such as ensuring sufficient deck space, feasibility of liquid transfers, and maximum and dead volumes for labware.

SynthaceHub automatically sends log and configuration files to the cloud for storage in your organization’s private data repository in Synthace from the liquid handler’s native software, such as Tecan’s Freedom EVOware® for the Freedom EVO®.

SynthaceHub uses a range of data ingestion methods depending on the hardware involved to send experimental data files to the cloud for storage in your organization’s private data repository in Synthace. If you would like to learn more about the technical details, please drop us an email or use the “Request a  demo” button below.

Yes, Synthace supports exports to appropriate file formats for further analysis with Excel, Python, and more.

Yes, Synthace's cherry picker is one example of that. Another example would be DOE runs with hard to change factors where we would need to run across two workflows.

Experimental results can be searched and accessed by everyone in your group/organization who has an account in Synthace. If you would like to discuss access control and data security in more detail, please drop us an email or use the “Request a demo” button below.

Although calendar integrations are not yet available in Synthace, the estimated run times as calculated by Synthace’s planning engine provide an accurate timing for you to ensure that you have booked enough time on your desired devices to execute your experiments.

Technical note

How can working with Synthace enhance your current RoboColumn® workflows?

Our technical note compares miniaturized purification using Tecan’s Freedom EVOware® Te-Chrom Wizard and Synthace's cloud-based Synthace.  

See how Synthace is able to increase flexibility and save time.

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