To begin, prepare the organic phase extraction solution and reservoir by adding the extraction solution to a two liter glass reservoir and placing it on a magnetic stir plate. Next, position a 100 RPM variable speed pump drive with a polytetrafluoroethylene diaphragm pump head next to the reservoir. Using flexible pump tubing, connect the outflow port of the extraction solution reservoir to the diaphragm pump inlet.
Then connect the pump outlet to the shell-side inlet at the base of the forward extraction membrane. Now, connect the shell-side outlet at the top of the forward extraction membrane to the shell-side inlet located at the base of the backward extraction membrane. Then connect the shell-side outlet at the top of the backward extraction membrane to the return port on the extraction solution reservoir.
To prepare the aqueous phase stripping solution and reservoir, pour the solution into a four liter glass reservoir with a magnetic stir bar and place it on a magnetic stir plate. Integrate a pH control system with a set point of pH nine with the stripping solution reservoir. Insert the pH probe through the stripping solution reservoir port and suspend it in the stripping solution.
To install the hollow fiber membrane filter, place a 300 RPM variable speed pump drive adjacent to the bioreactor. Affix the hydrophilic hollow fiber membrane to a ring stand above the peristaltic pump. Stack two peristaltic pump heads, large and small, on the pump drive.
Now using flexible tubing, connect the outflow port at the bioreactor to the large pump head inlet. Then connect the large pump head outlet to the hydrophilic hollow fiber membrane filter at the tube-side inlet located at the base of the filter. Cap the top shell-side port of the filter to prevent the influx of air.
To connect the forward extraction membrane, affix the first hydrophobic hollow fiber membrane module to a ring stand above the pump. Using flexible pump tubing, connect the hydrophilic hollow fiber membrane filter at the shell-side outlet to the small pump head inlet. Then connect the small pump head outlet to the forward extraction membrane at the tube-side inlet located at the base of the module.
Now using a coupling fitting and T fitting, connect the pressure gauge and flow restriction valve to the tube-side outlet of the forward extraction membrane. Next, connect the tube-side outlet of the forward extraction membrane to the return port at the bioreactor. To connect the backward extraction membrane, place a 300 RPM variable speed pump drive adjacent to the bioreactor.
Affix the second hydrophobic hollow fiber membrane module to a ring stand above the peristaltic pump. Then using flexible pump tubing, connect the stripping solution reservoir outflow port to the peristaltic pump inlet and the pump outlet to the tube-side inlet at the base of the backward extraction membrane. Afterward, using a coupling fitting and T fitting, connect the pressure gauge and flow restriction valve to the tube-side outlet of the backward extraction membrane.
Using flexible pump tubing, connect the tube-side outlet at the top of the backward extraction membrane to the stripping solution reservoir return flow port.