FOULING INDUCED ENZYME IMMOBILIZATION USING ULTRA FILTRATION MEMBRANE THROUGH TANGENTIAL FLOW FILTRATION: ADVANCES AND IMPLICATIONS IN BIO-PHARMACEUTICAL DOWNSTREAM PROCESSING

The biopharmaceutical industry is one of the most scientifically demanding fields in healthcare, focused on the production of therapeutic proteins, monoclonal antibodies (mAbs), nucleic acids, and other biologically derived agents. These products are typically generated using recombinant DNA technology, cell culture, or microbial fermentation. Due to their complexity and sensitivity, maintaining high product yield, purity, and integrity during downstream processing (DSP) is essential. However, membrane fouling, especially biofouling, remains a significant challenge in DSP operations such as tangential flow filtration (TFF), ultrafiltration (UF), and diafiltration (DF), impairing filterability and membrane longevity. To address this, enzyme immobilization has emerged as a promising strategy, particularly when integrated with membrane-assisted systems. Immobilizing enzymes onto or within membranes enhances enzyme stability, supports reusability, and enables coupling of catalytic and separation functions. A novel approach gaining attention is fouling-induced immobilisation, which utilises natural fouling phenomena, such as pore blockage, cake layer formation, and surface adsorption, to entrap enzymes without requiring chemical modification. TFF is especially well-suited for such applications due to its ability to regulate shear stress and control fouling dynamics. Innovative membrane configurations, such as the ―sandwich mode,‖ where a porous support is layered above the membrane, have demonstrated improved enzyme retention and sustained flux. The choice of membrane material also plays a key role: hydrophilic regenerated cellulose supports compatibility with enzymes, while polysulphone membranes promote adsorption, albeit with greater fouling risks. TFF thus serves as both a filtration and biocatalysis platform in modern bioprocessing.