Introduction:
Nowadays, through the cultivation of mammalian cells, a new generation of biopharmaceuticals has been developed as never before. This article mainly introduces the process of separating proteins secreted by mammals by hollow fiber micropore filtration. At an initial concentration of 2 × 10 5 cells/ml, the extracellular protein product is a 10kD lymphokine similar to interleukin-2 and is a potential tumor treatment drug. This application requires removal of cells and particles without lysis of cells while achieving high protein recovery. Processing time is critical because the product is thermally unstable.
KrosFlo hollow fiber filter assembly
Process conditions:
In view of the low cell concentration, a throughput of 600 L typically requires only a filter having a membrane surface area of ​​2-3 m 2 . However, this treatment required a shorter time, so two KrosFlo modules with a molecular weight cut-off of 0.2 μm were installed in parallel, each having a surface area of ​​3.3 m 2 . The cycle speed is expected to be 70 L/min. The filter inlet pressure is fixed at 5 psig. When the amount of filtration reached 30% of the throughput, a 10 psi back pressure was applied at the filter outlet, i.e., the filter inlet pressure was raised to about 15 psig. With the exception of minor adjustments, all conditions remain the same during processing.
result:
The 600L sample was processed for filtration at 65 min. The steady state flow rate was 85 L/m 2 hr. The amount of filtration is basically linear with time. To maximize filtration, the pressure and flow are slightly optimized during processing. As expected, the filter's flow rate decays slightly over time, but the processor's product recovery is still 94% or higher.
Flux & Filtration vs. Time
discuss:
In this treatment, the target protein will be tightly bound to the polysulfone membrane, but economically, a higher yield is required. The KrosFlo fiber membrane has a good protein channel and as a single-use component, it maintains consistent membrane properties from the start to the end of the process, eliminating the cost and time required for membrane cleaning and avoiding the risk of contamination. Spectrum's pure membrane modules are available in a variety of surface area specifications to facilitate process time optimization. The data shows that the use of microporous hollow fiber cross-flow filtration can simultaneously achieve rapid filtration and high recovery of heat sensitive products.
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