Automated Protein Purification and Buffer Exchange with IMCStips® and SizeX® tips on the CyBio FeliX

The CyBio FeliX is a compact benchtop automated liquid handler that has enough deck space to hold all the required consumables and reagents while remaining small enough to fit in a standard sized Biosafety hood. This gives you the flexibility to rapidly Purify and Buffer Exchange anywhere between 1 and 96 proteins without compromising on a sterile environment. The CyBio FeliX puts you in control, allowing you to increase or decrease the binding, washing and elution volumes and cycle numbers to optimise the purification for your sample type. 

IMCS offer CyBio FeliX 1000µl tips with variable resin volumes depending on the required yield of protein purified. The recommendations for most agarose resins (with binding capacities of 50-80 mg/ml) are:

• 10µl Resin Volume for 200ug or fewer

• 25µl Resin Volume for 200ug – 1mg

• 50µl Resin Volume for 1mg and above

The number of binding cycles is determined by several factors. These include, resin volume, sample volume, concentration of protein in sample, and desired purified protein concentration. Optimisation will be needed and these figures below are a guide

• Flexible sample numbers 1 - 96

• Full plate Purification and Buffer Exchange on a single deck

• Can handle sample volumes greater than 1ml

• Fits into a BSC or HEPA cabinet

When purifying 1-96 samples, the FeliX Deck is loaded as follows:

1. Removal of storage buffer

   The CyBio FeliX R96/1000 head aspirates before tip are loaded. The IMCS Resin Tips are loaded, and the storage buffer is forced out to the Waste Plate. The tips are then unloaded back to their Tip Position, and the R96/1000 Head aspirates before loading the IMCS SizeX Tips and forcing the storage buffer to the Waste Plate again.

    

2. Equilibration of Resin

   The IMCS Resin tips move to the Plate on deck loaded with Equilibration Buffer. The Tips aspirate and dispense 800µl of the Equilibration Buffer for several cycles to remove any trace of storage buffer 

    

3. Sample Binding

   The CyBio FeliX aspirates up to 900µl of the sample into the IMCStips, where the patented disperser promotes efficient mixing between the sample and resin to maximise protein binding before dispensing back into the plate. The aspirate dispense cycles and pipetting speeds are easily optimised within the CyBio FeliX Composer Software to maximise binding rate.

    

4. Sample Washing

   The CyBio FeliX aspirates and dispenses the IMCS Resin Tips in the first plate containing Wash Buffer before moving onto the second wash buffer plate. The washing steps remove any left-over contaminants and small molecules resulting in high purity samples

    

5. Sample Elution

   The desired protein is unbound to the resin by aspirating and dispensing elution buffer. The number of mixing cycles is optimised for maximum recovery for both Elution Plate 1 and 2. After completion, your proteins have been purified

    

6. Equilibration and sample addition to SizeX Tips

   The CyBio Felix 96/1000 tips are loaded to transfer 300µl SizeX Equilibration Buffer from the SizeX Equilibration and Chaser Plate to dispense inside of the SizeX Tips. The CyBio FeliX swaps to the SizeX Tips and forces the equilibration buffer into the waste plate. This is completed 3 times to remove any left-over storage buffer and equilibrates the tips. The sample is dispensed into the SizeX Tip, where the purified protein is captured within the hydrophilic polymeric resin.

    

7. Chaser and capture of protein

   The Chaser Buffer is dispensed into the SizeX Tip and then loaded into the R96/1000 CyBio FeliX Head. The Head moves above the Waste Plate and dispenses the unwanted salts and buffers, before forcing the purified and buffer exchanged proteins into the output plate.