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Un sistema totalmente automatizado de alto rendimiento para enriquecer y separar simultáneamente los glicopéptidos

Zhonghan Hu¹, Rong Liu², Junhui Li²

¹Institute of Mass Spectrometry, Zhejiang Engineering Research Center of Advanced Mass spectrometry and Clinical Application, Ningbo University, Ningbo 315211, PR China; Zhenhai Institute of Mass Spectrometry, Ningbo 315211, PR China; School of Marine science, Ningbo University, Ningbo 315211, PR China.

Journal of chromatography. A

|February 11, 2026

Resumen

Un nuevo sistema automatizado, AutoGP 2, mejora el análisis de glicopéptidos al permitir el enriquecimiento y la separación simultáneos. Esta herramienta de glicoproteómica de alto rendimiento mejora la eficiencia del análisis de muestras y la reproducibilidad para estudios clínicos.

Área de la Ciencia:

* Proteómica y Glicómica.
* Química analítica * Química analítica
* Automatización y sistemas de alto rendimiento.

Sus antecedentes:

* El análisis de glicopéptidos es crucial para comprender los procesos biológicos.
* Los métodos existentes para el enriquecimiento y la separación de glicopéptidos pueden consumir mucho tiempo y limitar el rendimiento.
* Hay una necesidad de soluciones automatizadas de alto rendimiento para estudios glicoproteómicos a gran escala.

Objetivo del estudio:

* Desarrollar un sistema mejorado y totalmente automatizado de alto rendimiento (AutoGP 2) para el enriquecimiento y separación simultáneos de glicopéptidos intactos.
* Para evaluar el rendimiento, incluido el rendimiento, la sensibilidad y la reproducibilidad del sistema AutoGP 2.
* Para demostrar la idoneidad del sistema para el análisis de muestras glicoproteómicas clínicas de cohorte grande.

Principales métodos:

* Introducción de una segunda válvula de 10 puertos en el sistema AutoGP existente para permitir el procesamiento simultáneo.
* Evaluación sistemática de los parámetros óptimos de funcionamiento para el sistema AutoGP 2.
* Evaluación de la reproducibilidad intradiaria e interdiaria mediante pruebas de esfuerzo de 24 horas y 15 días.
* Acoplamiento de AutoGP 2 con un espectrómetro de masas Orbitrap de alta resolución.

Principales resultados:

* AutoGP 2 mejora significativamente el rendimiento del análisis de muestras en comparación con el sistema AutoGP anterior.
* Se identificaron más de 560 glicanos específicos del sitio a partir de una mezcla de péptidos de 1 μg con una CV media del 10,8% en análisis triplicados.
* El ciclo de trabajo del instrumento aumentó del 55% al 80%, ahorrando 166 minutos por análisis de dos muestras.
* Demostró una sensibilidad y reproducibilidad sobresalientes.

Conclusiones:

* AutoGP 2 es un sistema automatizado altamente eficaz para el enriquecimiento y la separación simultáneos de glicopéptidos intactos.
* El sistema ofrece mejoras significativas en el rendimiento, la sensibilidad y la reproducibilidad para la glicoproteómica.
* AutoGP 2 es muy adecuado para el análisis de muestras glicoproteómicas clínicas a gran escala y estudios limitados a muestras.

Palabras clave:

El enriquecimiento automatizado.La glucosilación por glicosilación Alta reproducibilidad con alta reproducibilidad.Alto rendimiento de alto rendimiento.El glicopéptido intacto.

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