Automated Protein purification or Plasma DNA Workflow with PhyTip technology
Proteomics is currently one of the most important scientific fields with the growing DNA sequence information. It provides insights in biological processes and their mechanisms. With the PhyTip technology automated on the Dynamic Devices Lynx liquid handling platforms it is possible to examine the expression of over 1.000 proteins in high throughput using mass spectrometry coupled to separation techniques.
Dynamic Devices Lynx platforms
The Dynamic Devices Lynx system is a liquid handling platform available in various dimensions. The smallest platform (Lynx LM700) is only 70 cm in deck space, but still accommodates 9 or 12 deck positions. The most used Lynx LM900 offers 30 deck plate positions in a deck of just 90 cm wide. For increased throughput the Lynx LM1200 and Lynx LM1800 offer 42 and 66 microplate positions and can be equipped with two pipetting arms. For the highest throughput possible, the Lynx LM730i is a system specifically designed for integration in a completely automated setting.
The Dynamic Devices Verified Volume Pipetting technique
With the Verified Volume Pipetting technology, Dynamic Devices sets itself apart from any other liquid handling platform available in the market today. With the Volume Verified Pipetting technique you have full control over all 96 channels in the head. With the 96 fully independent channels, there is no need any more for an 8 channel arm. Due to the flow sensor technology, the system is able to aspirate and dispense all fluids no matter what the viscosity is and the technique makes liquid classes a thing of the past. The VVP technology provides full tracking and tracing of each and every single channel through every liquid transfer including clot detection. The VVP technology in combination with the Phynexus PhyTip separation solutions accommodate for the highest control and throughput during the automated protein purification or plasma DNA workflow.
Real Time Transfer Validation
Being awarded with the 2012 SLAS New Product Award, Dynamic Devices shows that with the Verified Volume Pipetting (VVP) they have your samples as their number 1 priority. The VVP technology enables real time transfer validation throughout your assay. When transferring 96 different volumes, the software shows full transfer validation.
Figure 1: Transfer Validation
Using the 96 VVP pipetting tool with clot/clog detection active, challenging samples are aspirated with individual channel diagnostics. If a clog/clot starts to interfere with the liquid aspiration, that individual tip is halted until the rest of the tips finish and then the system will clear and re-aspirate the erred channel. Especially in the Phynexus Phytip applications, the clot/clog detection, gives more control over your assay.
Figure 2: Clot/Clog Detection
Creating liquid classes is a thing of the past. With the Dynamic Devices VVP technology it is no longer required to preset for various liquid types. The VVP directly measures the flow rate of the liquid in the tip and accommodates for viscosity or temperature. The only thing needed is a target volume and the VVP technology and valves make sure the correct volume is aspirated in the tip.
Phynexus Phytip chromatography technology
Phynexus Phytip technology is a patented chromatography tip for high throughput affinity chromatography for protein and plasma purification. The PhyTip column technology is the most efficient and cost effective way to purify recombinant proteins, antibodies and plasmids. The technology allows for bidirectional flow and elution volumes as small as 10 μl.
Figure 3: Phynexus PhyTip technology
Dynamic Devices and PhyTip technology
The Dynamic Devices Lynx designed for PhyTip technology provides the following accessories:
- PhyTip 1000 μl Tip Station
- PhyTip Drying Station
- Standard 1000 μl Tips
- Deepwell sample plates
- Elution plates
- Column Prep Reagent reservoir
- Deepwell Wash Buffer 1
- Deepwell Was Buffer 2
- Elution buffer reservoir
The Deepwell plates for buffers are used to prevent cross-contamination between samples during the run. The Wash Buffers are transferred into the deepwell plates from a reservoir into the plates and removed after processing.
Processing 1-96 samples processing in parallel
To create a complete walk-away solution, Dynamic Devices offers the Lynx LM 900 with 30 plate positions in combination with the PhyTip technology. It is a bench-top solution with a high level of control and precision for the preparation of protein and plasmids samples. It allows for the unattended parallel processing of a range of samples by utilizing the industry standard resins. Columns can be used to process samples from 10 μl up to 25 ml. If the tip volume is exceeded, the samples are processed as a series of fractions throughout the run.
To avoid impurities, the leading and trailing edge of the peak are not collected, only the center of the peak will be collected through the heart cut process. Normally this is a tedious and relatively expensive method to operate in parallel. Within the automated setup, a virtual heart cut method is applied for superior protein purity and concentration. The process is started by fully loading to capacity of the pipette tip column with the resins. This is achieved by back-and-forth flow of the sample through the PhyTip column. This process assures that even the slow interaction proteins have sufficient contact time to interact with the resin.
The following step is the complete washing of the resin bed with a very high wash to resin volume to assure a high purity. Washing can be done up to 50 times and will remove nonspecific bound contaminants. To perform the wash, the thin, low dead volume and low surface area column frits are used.
To produce the highest concentration possible, the process allows for fine control of the elution volume. Due to the high loading of the column and the small elution volume. The virtual heart-cut Elution process produces a final product comparable to the recovery of the heart cut technology. The final result therefor is a 5-10 times higher concentration compared to other technologies.