Preparative scale production and functional reconstitution of a human aquaglyceroporin (AQP3) using a cell free expression system.

•We establish an efficient cell-free strategy to produce AQP3, an aquaglyceroporin.•Milligrams quantities of high quality AQP3 are made in a single reaction, in ml volumes.•The developed cell-free system is reliable, easily scalable and flexible.•Other aquaglyceroporins could be made at preparative scale by the devised strategy.•The proposed system may accelerate high-throughput studies and translational studies.