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 single-use flexible container with an inner volume and capable of holding nutrient media;
• ii) the single-use flexible container of (a)(i) having at least one interior wall, 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 with a variable density over the surface of the septum, wherein diameter of the pores range in size from 1 μm to 1000 μm, and said septum provides fluid communication between the right and the left chambers;
• 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 capable 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 heater and/or a cooling element for heating and/or cooling the flexible container;
• xi) a motor driving a shaft connected to a rolling mechanism for pivoting the cylindrical support along its circular axis between −110 to +110 degrees;
• 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 with 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) moving the shaft to pivot the cylindrical support between the angles from −110 and +110 degrees 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.