E. coli lysate clarified, harvesting intracellular expressed protein

Introduction

The proteins expressed in the cells were separated from the E. coli lysate. The protein of interest has anti-tumor potential.

Process conditions

The lysate volume was 245 liters. Batch separation was performed using a 0.2 μm Krosflo ^® module with a membrane surface area of ​​1.0 m ² . The flow path is shown in Figure 1, and the circulation rate of the feed liquid is maintained at 57 L/min. Start the feed cycle first, then slowly open the filtrate port. The downstream loop is not limited. The initial processing temperature was 4 ° C and slowly rose to 12 ° C until the end of the run. The initial injection pressure was 12 psig, as the temperature increased, the viscosity of the feed dropped, the pressure dropped to 11 psig, and then increased to 13 psig due to the increase in solids concentration before the end of the run.

Figure 1. E. coli lysate filtration flow path system setup

Figure 2. E. coli lysate filtration flux and total filtrate volume vs. time

result

As a typical bacterial lysate, the flow decay curve begins to decrease rapidly and quickly stabilizes. A total of 237 L of the filtrate containing the protein of interest was collected in 65 min. The average throughput of the process was ~200 L/m ² /hr, and the pass rate of the target protein was 99%.

discuss

Selection of suitable filtration components and process conditions is necessary to effectively separate the desired component of interest from the cell lysate. The Spectrum Pure Krosflo® Filter Kit is the best choice for separating soluble proteins from cell lysates. If the disposable pre-sterilization filter is used for treatment, the problem of insufficient filter cleaning can be avoided. Filters no longer cause contamination between batches and guarantee high efficiency for each treatment.

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