Filtration and separation technologies remain as one of the biggest challenges humanity currently faces. The separation of different elements such as bacteria, viruses, heavy metals, particles, and chemical agents require the development of multifunctional membranes. In membrane technology, one of the most promising fabrication techniques is electrospinning, which can produce highly tailored non-woven fibrous multifunctional membranes with a high surface area. On the other hand, cellulose derivatives, like cellulose acetate, have many beneficial properties for filtering technology such as high availability and easy functionalization. Likewise, cellulose nanocrystals are used to improve mechanical properties and functionalize membranes. In this project, a cellulose nanocrystal (CNCs) functionalized cellulose acetate electrospun multifunctional membrane is developed for adsorption and separation of nanosized particles. In this work, cellulose acetate (CA) fibers with an average fiber diameter of approximately 900 nm were electrospun and tested as membranes for size and affinity based filtration. First, the electrospinning process was optimized regarding solution and process parameters. As a result, solution parameters were found to be 12 wt% solid content CA dissolved in a 1:1 acetone:acetic acid solution. Regarding process parameters, the suitable electrospinning parameters were found to be 18 kV applied voltage, a feeding rate of 5 mL/h, and a tip-to-collector distance of 20 cm.   The electrospun CA membrane was coated with cationic (+) and anionic (-) cellulose nanocrystals up to a 25 wt% concentration. The incorporation of CNCs, of either anionic or cationic surface charge, affected membrane wettability. The neat CA membrane had a hydrophobic behavior with a contact angle of 110°. The addition of CNCs decrease contact angle, to 31.5° for CA-CNCs(-) and 50° for CA-CNCs(+), which resulted in functionalized membranes with a hydrophilic behavior. Both functionalized membranes managed to maintain high flux values. CA-CNCs(-) maintained a flux of 9500 Lm−2h−1, while CA-CNCs(+) maintained a flux of 6700 Lm−2h−1. The addition of cellulose nanocrystals improved the mechanical properties of the CA membranes. The tensile strength increases from 410 kPa to 4990 kPa for CA-CNCs(-) membranes and 3010 MPa for CA-CNCs(+) membranes, and is accompanied by an increase in Young’s modulus as well. To evaluate the adsorption efficiency and size-exclusion filtration, an anionic dye (Congo red), a cationic dye (Victoria blue), and 500 nm model particles were used. CA-CNC(-) membranes achieved a removal efficiency of 96% of 500 nm particles with an affinity-based dye removal of 63% of Victoria blue dye. On the other hand, CA-CNC(+) membranes achieved a removal efficiency of 43% of 500 nm particles with a dye removal of 27% of Congo red dye. In this regard, CA-CNC(-) membranes were the best candidate for size-exclusion filtration, while also maintaining a good level of adsorption. Cellulose based composite membranes were successfully produced as multifunctional filters that could act in both size-exclusion regime and affinity-based regime. A cellulose acetate fibrous membrane was produced by electrospinning, tuning for fiber size and porosity, while the incorporation of cellulose nanocrystals functionalizes the membranes and enhance mechanical properties, and wettability.