Selección, expresión recombinante y caracterización de nanoanticuerpos de tipo camélido bloqueadores de TNF-α humano.
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Date
2025
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Universidad de Concepción
Abstract
La autoinmunidad es una condición del sistema inmunológico caracterizada por la generación de moléculas autorreactivas. Si bien su presencia está regulada por mecanismos de tolerancia inmunológica que permiten la eliminación de células autorreactivas y la modificación de proteínas de membrana, estos procesos pueden fallar, conduciendo al desarrollo de enfermedades autoinmunes. Tales patologías, incluidas las enfermedades inflamatorias intestinales como la colitis ulcerosa y la enfermedad de Crohn, así como la psoriasis y la artritis reumatoide, afectan entre el 3 % y el 5 % de la población mundial. En particular, la artritis reumatoide, que impacta aproximadamente al 1 % de la población global, se caracteriza por provocar un daño crónico severo al sistema musculoesquelético.
El factor de necrosis tumoral alfa (TNF-α), una citocina homotrimérica de 157 aminoácidos expresada principalmente por macrófagos activados, desempeña un papel central en la respuesta inflamatoria y está directamente implicado en la fisiopatología de la artritis reumatoide. Actualmente, cinco fármacos aprobados por la Administración de Alimentos y Medicamentos de los Estados Unidos (FDA) emplean estructuras de anticuerpos completos, fragmentos de anticuerpos o proteínas de fusión dirigidas contra TNF-α o sus receptores, producidos en sistemas bacterianos o en células de mamífero, siendo estas últimas las plataformas más utilizadas. No obstante, ambos sistemas presentan altos costos de producción, lo que incrementa significativamente el precio final del tratamiento.
En este contexto, el presente trabajo tiene como objetivo expresar y caracterizar nanoanticuerpos de tipo camélido con capacidad de bloquear TNF α. Se propone el uso de una nueva molécula con potencial terapéutico y la optimización de procesos biotecnológicos orientados al desarrollo de bioproductos innovadores, con el fin de ofrecer alternativas terapéuticas más accesibles y eficientes para el tratamiento de enfermedades autoinmunes.
Autoimmunity is a condition of the immune system characterized by the generation of self-reactive molecules. Although their presence is regulated by immune tolerance mechanisms that allow for the elimination of self-reactive cells and the modification of membrane proteins, these processes can fail, leading to the development of autoimmune diseases. Such pathologies, including inflammatory bowel diseases such as ulcerative colitis and Crohn's disease, as well as psoriasis and rheumatoid arthritis, affect between 3% and 5% of the global population. In particular, rheumatoid arthritis, which affects approximately 1% of the global population, is characterized by causing severe chronic damage to the musculoskeletal system. Tumor necrosis factor alpha (TNF-α), a homotrimeric cytokine composed of 157 amino acids, mainly expressed by activated macrophages, plays a central role in the inflammatory response and is directly implicated in the pathophysiology of rheumatoid arthritis. Currently, five drugs approved by the United States Food and Drug Administration (FDA) employ full antibody structures, antibody fragments, or fusion proteins directed against TNF-α or its receptors, produced in bacterial systems or mammalian cells, the latter being the most commonly used platforms. However, both systems present high production costs, which significantly increase the final price of the treatment. In this context, the present work aims to express and characterize camelid-type nanobodies capable of blocking TNF-α. The use of a new molecule with therapeutic potential and the optimization of biotechnological processes aimed at the development of innovative bioproducts is proposed, with the objective of offering more accessible and efficient therapeutic alternatives for the treatment of autoimmune diseases.
Autoimmunity is a condition of the immune system characterized by the generation of self-reactive molecules. Although their presence is regulated by immune tolerance mechanisms that allow for the elimination of self-reactive cells and the modification of membrane proteins, these processes can fail, leading to the development of autoimmune diseases. Such pathologies, including inflammatory bowel diseases such as ulcerative colitis and Crohn's disease, as well as psoriasis and rheumatoid arthritis, affect between 3% and 5% of the global population. In particular, rheumatoid arthritis, which affects approximately 1% of the global population, is characterized by causing severe chronic damage to the musculoskeletal system. Tumor necrosis factor alpha (TNF-α), a homotrimeric cytokine composed of 157 amino acids, mainly expressed by activated macrophages, plays a central role in the inflammatory response and is directly implicated in the pathophysiology of rheumatoid arthritis. Currently, five drugs approved by the United States Food and Drug Administration (FDA) employ full antibody structures, antibody fragments, or fusion proteins directed against TNF-α or its receptors, produced in bacterial systems or mammalian cells, the latter being the most commonly used platforms. However, both systems present high production costs, which significantly increase the final price of the treatment. In this context, the present work aims to express and characterize camelid-type nanobodies capable of blocking TNF-α. The use of a new molecule with therapeutic potential and the optimization of biotechnological processes aimed at the development of innovative bioproducts is proposed, with the objective of offering more accessible and efficient therapeutic alternatives for the treatment of autoimmune diseases.
Description
Tesis presentada para optar al grado de Doctor en Biotecnología Molecular.
Keywords
Anticuerpos monoclonales, Proteínas recombinantes, Inmunología molecular