Pivoting pressurized single-use bioreactor

A method for producing a biological product comprising:

• a) Providing a bioreactor including:
• i) a cylindrical support with an inner surface configured to immovably retain a flexible container with a inner volume and capable of holding nutrient media;
• ii) A single-use flexible container having at least one interior wall and an inner volume, further including:
• A flexible septum with a surface immovably positioned within the flexible container and defining a right chamber and a left chamber, the septum having a plurality of pores ranging in size from 1 μm to 1000 μm that provides fluid communication between the right and the left chambers and the surface of septum having variable density of the plurality of pores;
• iii) Disposing the flexible container immovably inside the cylinder such that the septum is vertically positioned and a surface comprising 25-30% of the total surface of septum along the middle of the horizontal axis and the middle of the vertical axis has 3-5 times higher density of pores than the rest of the septum.
• iv) At least one liquid inlet;
• v) At least one liquid outlet;
• vi) At least one gas inlet in fluid communication with a source of compressed gas further comprising a sterilizing filter positioned between the source of compressed gas and the container;
• vii) At least one gas outlet with means of controlling the rate of flow of gas;
• viii) at least one sensor to measure the pressure inside the flexible container;
• ix) at least one sensor to measure the temperature of the liquids in the flexible container;
• x) A means of heating or cooling the flexible container;
• xi) a means of pivoting the cylindrical support along its circular axis between −110 to +110 degrees;
• xii) an electronic means of controlling the frequency and the degree of pivoting of the cylinder, the pressure in the container and the temperature of the nutrient media in the flexible container.
• b) Disposing the flexible container in the cylindrical support and immovably fixing the flexible container to the inner surface of the cylindrical support;
• c) Introducing the nutrient media in the flexible container through the liquid inlet;
• d) Introducing a biological culture capable of growing in the nutrient media and producing a biological product in the flexible container through the liquid inlet;
• e) Heating or cooling the nutrient media to a pre-determined temperature;
• f) Connecting the gas inlet to a source of a compressed gas;
• g) Starting the flow of the compressed gas with sufficient pressure to achieve a pre-determined pressure in the flexible container that causes the flexible container to expand and stay in continuous contact the inner surface of the cylindrical support;
• h) Continuing the flow of compressed gas to maintain the pre-determined pressure in the flexible container;
• i) Starting the means of pivoting the cylindrical support to pivot it between the angles from −110 and +110 at a pre-determined frequency;
• j) Monitoring the density and the viability of the biological culture in the nutrient media at predetermined time intervals;
• k) Monitoring the concentration of the biological product in the nutrient media at predetermined time intervals;
• l) Stopping the pivoting of the cylindrical support when the density of the biological culture or the concentration of the biological product reaches a pre-determined level and removing the nutrient media from the flexible container for further processing of the purification of the biological product in the nutrient media.