Valente Joana F A, Carreira Tiago Soares, Dias Juliana R, Sousa Fani, Alves Nuno
CDRSP-PL-Centre for Rapid and Sustainable Product Development, Polytechnic of Leiria, 2411-901 Leiria, Portugal.
CICS-UBI-Health Science Research Centre, University of Beira Interior, Av. Infante D. Henrique, 6200-506 Covilha, Portugal.
Pharmaceutics. 2022 Oct 23;14(11):2266. doi: 10.3390/pharmaceutics14112266.
The increasing progression of biopharmaceutical-based therapies highlights the demand for efficient chromatographic methods that can be used to purify the desired biomolecules (e.g., nucleic acids, enzymes, or monoclonal antibodies) which are presently under consideration in clinical trials or approved by the Food and Drug Administration. These molecules present distinct chemical and structural properties, which are critical cues for the development and production of adequate chromatographic supports. Until now, it has not been possible to fully control the characteristics of the chromatographic matrices to assure the total reproducibility of their structure and packing. Meanwhile, three-dimensional printing (3DP) is in the early stage of its use in the production of chromatographic supports as a fast, very precise, and reproducible methodology. Although 3DP can provide excellent performance properties to the chromatographic structures, it cannot, per se, lead to high-quality pharmaceutical products. However, the association of affinity ligands, such as amino acids, which is possible in 3DP, could enable the attainment of high-purity yields of the desired molecules. Beyond the amino acids most widely studied as chromatographic ligands, arginine has been successfully immobilized on different chromatographic supports (namely, agarose bead matrices, macroporous matrices, and monoliths) to achieve extra-pure gene therapy products. In this research, we studied the immobilization of arginine on 3DP chromatographic supports, evaluating the stability of the ligand/chromatographic support linkage under different chromatographic conditions to determine the robustness of these new prototypes. Moreover, we also applied plasmid DNA samples to these supports to observe the practical behaviour of the developed arginine 3DP chromatographic structures.
基于生物制药的疗法的不断发展凸显了对高效色谱方法的需求，这些方法可用于纯化目前正在临床试验中或已获美国食品药品监督管理局批准的所需生物分子（例如核酸、酶或单克隆抗体）。这些分子具有独特的化学和结构特性，这是开发和生产合适色谱支持物的关键线索。到目前为止，还无法完全控制色谱基质的特性以确保其结构和填充的完全可重复性。同时，三维打印（3DP）作为一种快速、非常精确且可重复的方法，在色谱支持物生产中的应用尚处于早期阶段。尽管3DP可为色谱结构提供优异的性能，但就其本身而言，它无法生产出高质量的药品。然而，在3DP中可以实现亲和配体（如氨基酸）的结合，这可能使所需分子获得高纯度产率。除了作为色谱配体被广泛研究的氨基酸外，精氨酸已成功固定在不同的色谱支持物上（即琼脂糖珠基质、大孔基质和整体柱），以获得超纯的基因治疗产品。在本研究中，我们研究了精氨酸在3DP色谱支持物上的固定化，评估了配体/色谱支持物连接在不同色谱条件下的稳定性，以确定这些新原型的稳健性。此外，我们还将质粒DNA样品应用于这些支持物，以观察所开发的精氨酸3DP色谱结构的实际性能。
