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Exploración de la estabilidad de las células CHO durante el pase prolongado a través del análisis de balance de flujo

Dong-Hyuk Choi1, Sun-Jong Kim1, Jinsung Song1

  • 1School of Chemical Engineering, Sungkyunkwan University, Suwon, Gyeonggi-do, Republic of Korea.

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PubMed
Resumen

El cultivo a largo plazo de células de ovario de hámster chino (CHO) para la producción de proteínas enfrenta desafíos de estabilidad. Este estudio revela que las células CHO de pases tardíos cambian su metabolismo del crecimiento a la gestión del estrés oxidativo, lo que afecta los rendimientos de proteínas terapéuticas.

Palabras clave:
células CHOestabilidad de producciónpase celularmetabolismo celularproteínas terapéuticasanálisis de balance de flujoIA explicableestrés oxidativotasa de crecimientotítulos de IgG

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Área de la Ciencia:

  • Biotecnología; Biología Celular; Ingeniería Metabólica

Sus antecedentes:

  • La estabilidad de la producción en líneas celulares de ovario de hámster chino (CHO) es crucial para la fabricación de proteínas terapéuticas, pero a menudo se ve comprometida durante el pase celular extendido.
  • Los mecanismos subyacentes que impulsan la inestabilidad y la reducción de la productividad en las culturas de CHO de pase tardío siguen siendo poco comprendidos.

Objetivo del estudio:

  • Caracterizar mecanicísticamente las diferencias fenotípicas y metabólicas entre las culturas de CHO de pase temprano (EP) y de pase tardío (LP).
  • Identificar las vías metabólicas clave responsables de la divergencia en el comportamiento y la productividad celular durante el pase extendido.

Principales métodos:

  • Análisis multivariante de datos (MVDA) de perfiles exometabolitos temporales.
  • Análisis de balance de flujo (FBA) integrado con inteligencia artificial explicable (xAI) para interrogar la reconfiguración metabólica.
  • Comparación del crecimiento celular, los títulos de proteínas terapéuticas (IgG) y las concentraciones de metabolitos clave entre las culturas EP y LP.

Principales resultados:

  • Las culturas de CHO de pase tardío (LP) mostraron densidades celulares viables máximas comparables, pero una reducción significativa (aproximadamente 35%) en los títulos máximos de IgG en comparación con las culturas de pase temprano (EP).
  • Las culturas LP exhibieron una mayor acumulación de lactato y amoníaco, lo que indica una función metabólica alterada.
  • El MVDA identificó la fase de crecimiento exponencial como la ventana crítica para la divergencia metabólica entre las culturas EP y LP.

Conclusiones:

  • Las células CHO de pase temprano (EP) priorizan el acetil-CoA para la biosíntesis de ácidos grasos para apoyar la proliferación.
  • Las células CHO de pase tardío (LP) cambian el enfoque metabólico hacia la mitigación del estrés oxidativo a través de la vía de la transsulfuración (síntesis de cisteína y glutatión) y una mayor actividad del ciclo TCA para la homeostasis energética.
  • El eje cisteína-glutatión representa un objetivo metabólico crítico para mejorar la estabilidad y la productividad a largo plazo de las culturas de células CHO.