Amaxa drives high-throughput cell transfection
consortium of companies, led by Amaxa, to develop the first ultra
high-throughput (UHT) devices for transfecting cells.
Transfection technology expert Amaxa will lead the consortium in its quest to develop UHT devices that can effectively deliver genetic material such as DNA or siRNA (small interfering RNA) into primary cells using Amaxa's Nucleofector technology before allowing subsequent cell screening in third party equipment. The European Commission (EC) has provided the consortium of eight European firms with a grant of €2.75m to aid the 36 month project. According to Amaxa, gene transfer technologies are crucial tools for studying gene regulation and gene silencing as well as for altering cells so that they produce biological drugs. However, Axama claims that the use of standard cell lines which are easily transfected can often give results of low biological and medical significance. Amaxa has developed a transfection technique based on electroporation which opens up pores into the cells through which biological material enters the cells. "You combine the cells and the target molecules with specific buffers in an electrical field and apply a high voltage for a few milliseconds and this drives the siRNA or DNA through the cell membrane and into the cells," said Dr Claus-Dietmar Pein, Amaxa's marketing director. "We have about 400 different protocols for different cell lines and you have to choose the correct conditions [buffer, voltage, time etc] otherwise you kill all the cells, if you choose the correct conditions you get up to 90 per cent survival." The technique can be used with primary cells and cell lines that are hard to transfect with the electroporation conditions being specific to the cells and not the biological material making the technique ideal for UHT screening of cells with a wide variety of biological agents. "The protocol is very cell specific but the target molecule that you want to get into the cell seems to have very little effect on our technology - there are other technologies on the market that can do gene transfer, but you would normally have to vary the conditions between different targets," said Dr Pein. He continued by saying that rather than just using the technology in drug discovery it could also find application in locating the best cell lines or genes for producing biological drugs. The new system will be designed to use 384 well plates that will minimise the size of the individual wells, leading to a reduction in the amount of material needed - potentially reducing the cost of each reaction by 75 per cent. The company clams that the method is significantly quicker than other methods and the expression of the transfected genes can be analysed after 2-4 hours rather than the 24-48 hours needed when using standard techniques. Because the Nucleofector technology is so much quicker than standard transfection techniques it makes an ideal basis for incorporation into automated high-throughput screening systems. "Amaxa is the only institution worldwide, which has the necessary knowledge and experience to adapt the technical needs of device development to the conditions used for efficient transfection of primary cells and hard-to-transfect cell lines," said Dr. Birgit Nelsen-Salz, coordinator of the project. "As coordinator of the project we will ensure that the expertise brought by each partner will enable the collaboration to achieve the goal of providing powerful new tools for basic research and drug discovery". Each partner in the collaboration, dubbed MODEST-EU, will have a significant role in device development, with Prevas (Sweden) responsible for device construction from proof-of-principle to the prototype stage while Fotec (Austria) will develop the micro-plate production process and HTP (Austria) will prototype the new 384-well plates. The nucleofection and cell based assays will then be assessed by: German cell sorting and multiparameter flow cytometry company DRFZ; German functional genomics RNAi technology company RNAx; Estonian central nervous system (CNS) and transcription experts, Protobios; and liver metastasis biomarker company Dominion Pharmakine.
