Producing Protein

Using its liquid handling robots – MPPU can process up to 96 samples or 96 different conditions per protein when expressed in E.coli, within 24 – 48 hours.

First published: 23/09/2014

Updated at: 24/09/2014 at 9:55 am

If your need is for large quantities of pure and active proteins, the Monash Protein Production Unit (MPPU) may have the solution.

MPPU is open for business and this is not just restricted to researchers at Monash.  The Protein Production Unit currently specialises in making recombinant proteins for research using expression services in any of four types of cell – E. coli, yeast, insect cell, and mammalian cells.

The amount of material generated is very dependent on the expression level of the specific protein of interest and the cell type used to express the recombinant proteins.

Using the latest high-throughput platforms for optimising protein expression and purification, the facility can produce pure and active proteins for use in research and development type projects.

Facility Manager Dr Noelene Quinsey said “Our process is automated with the latest high-throughput platforms. This makes it faster and more cost-effective than conventional manual methods.”

Using its liquid handling robots – PPU can process up to 96 samples or 96 different conditions per protein when expressed in E.coli. The processes are fast and cost effective to evaluate whether expression of the desired protein is possible in the E.coli system.

“Depending on what our customers need and whether the cell line given can be adapted to high throughput system, we can express a protein inside the cell membrane of E. coli within 24 – 48 hours.

Generally speaking, the PPU facility can produce

  • 20 litres of biomass for coli
  • 20 litres of biomass from yeast
  • 5-10 litres of insect cells, and
  • Presently up to 1.5 litres for transient mammalian cells (CHO and HEK293). We are currently testing scale up to 5L for the expression of a secreted and membrane proteins.

Prior to taking up the Facility Manger post at MPPU seven years ago, Noelene studied her post doctorate in expressing proteases in E. coli and mammalian cells, with Professor Rob Pike, focussing on understanding the biology of proteases involved in the blood clotting cascade in humans to enable development of compounds to protect against diseases.

It’s this kind of expertise within the facility that places MPPU at the forefront of protein expression and enables researchers to choose and replicate the conditions that give the best result.

“Different researchers use various expression vectors and have preferences for the cell system they want to work with. For example, E. coli can express recombinant proteins in an insoluble form which get isolated into a specialised compartment of the cell called the inclusion bodies.

“Proteins sent to the inclusion bodies are often mis-folded. But if there is a way to refold these recombinant proteins from the inclusion bodies, this can be a valuable source of starting material for the expression and purification of novel proteins.

“We have used this approach in the past to generate antigens for vaccine trials with great success. The trick is to know or understand what the function of the protein is when you refolded it to validate whether your refolded protein has a structure close to the native protein of interest.

“Researchers know what protein they want, but often don’t know where to start. I often start by researching the available literature myself, so I know the latest findings and procedures that are relevant to the protein of interest.” Dr Quinsey said.

Whether your research is in drug discovery, vaccine trials, structural biology, chemistry, protein therapeutics or biomedical imaging, the Protein Production Unit at Monash University can assist; offering Protein purification (up to 48 individual proteins), Custom purification (up to 100mg of protein on request), Purification Protocol development, Protein Expression Optimisation and Refolding screens/thermal melt analysis.

The facility’s equipment includes

  • TECAN evo 200 liquid handling robot
  • TECAN freedom evo liquid handling robot with 200 hood
  • AKTA Express units (12 Units)
  • RT-PCR machine
  • Bacterial/yeast/ insect cell growth facilities
  • Bioanalyser protein analysis
  • AKTA avant express unit
  • CO2 shaker incubators for the growth of mammalian cells in suspension culture.
  • Blitz biosensor

For more information click here 
or here or phone Dr Noelene Quinsey on +61 3 9902 0020.

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