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- Production of chromatographic supports by 3D printingPublication . Cravo, Priscilla Alice Ferreira; Valente, Joana Filipa Abreu Pereira; Sousa, Fani Pereira de; Alves, Nuno Manuel FernandesThe increased use of biopharmaceutical-based treatments shows the need for the development of efficient chromatographic techniques to purify specific biomolecules such as nucleic acids, enzymes, or monoclonal antibodies. These biomolecules are increasingly being employed in clinical trials and there are also several examples of approved biodrugs by the Food and Drug Administration (FDA). The unique chemical and structural characteristics that these biomolecules possess are crucial for developing suitable chromatographic materials. Until now, achieving complete control over the properties of chromatographic matrices and ensuring their consistent structure and packing has been a challenge. As a result, the slightly varied internal structure and porous configuration pose challenges in anticipating their chromatographic performance. This obligates to rigorous testing and validation of the packed chromatographic supports to ensure their quality before utilization. In this context, three-dimensional (3D) printing has been gaining recognition as a set of promising technologies for the production of chromatographic supports, with numerous advantages compared to traditional manufacturing methods. Among these advantages, there is the total control over the geometry of the produced piece, the design flexibility, one-step production, and the ability to easily conduct computational simulations on the desired geometry, such as simulating chromatographic runs (as the object is printed based on a computer model). Depending on specific requirements, various 3D printing processes can be employed, offering a variety of surface qualities and high dimensional precision. Although 3D printing can enhance the performance of chromatographic structures in terms of geometric standardization and efficient flow, these properties alone cannot guarantee the production of high-quality biopharmaceutical products. Therefore, it is necessary to modify them with ligands that ensure specific binding between the chromatographic matrix and the target molecule, ensuring high levels of purity and recovery of this molecule. Thus, in this work, Tyrosine was used as an affinity ligand. It possesses an aromatic ring and a hydroxyl group, making it suitable for participating in interactions in affinity chromatography, potentially serving to capture and purify biomolecules endowed with precise binding attributes. These biomolecules may include antibodies, enzymes, or DNA, and can selectively interact with tyrosine ligands. Within affinity chromatography, tyrosine ligands engage in intricate encounters with target molecules. These interactions unfold through a complex interplay of hydrophobic forces, hydrogen bonding, and other noncovalent interactions. These interactions rely on the distinct affinity that target molecules have for tyrosine, ultimately enabling the process of meticulous and selective binding, followed by purification. With this in mind, 3D printing was employed in this study to produce chromatographic supports with reproducible geometry, which were subsequently functionalized with Tyrosine. The experiments conducted in this study can be divided into four tasks: (1) Production of the 3D Printed chromatographic structure; (2) Tyrosine immobilization on the chromatographic support; (3) Assessment of the stability of the bonding between the ligand and the chromatographic support when subjected to different chromatographic conditions to assess the robustness of these new prototypes; (4) Binding experiments conducted with pDNA samples to understand the interaction profile of the developed matrices with this target molecule.