Sequential Injection Amperometric System Coupling with Bioreactor for In-Line Glucose Monitoring in Cell Culture Application

Abstract

We proposed a specially designed sequential injection (SI) amperometric system coupling with a bioreactor for in-line glucose monitoring in cell culture. The system is composed of three main parts which are the bioreactor, SI system, and electrochemical detection unit. The bioreactor accommodates six individual cell culture units which can be operated separately under different conditions. The SI system enables automatic in-line sampling and in-line sample dilution, with a specially designed mixing unit; therefore, it has the benefits of fast analysis time and less contamination risk. The use of 3D-printed microfluidic components, a mixing channel, and a flow cell helped to reduce operational time and sample volume. A disposable screen-printed electrode (SPE), modified with glucose oxidase (GOD), carbon nanotube, and gold nanoparticle, was used for detection. The developed system provided a linear range up to 3.8 mM glucose in cell culture media. In order to work with cell culture in higher glucose media, the in-line sample dilution can be applied. The developed SI system was demonstrated with mouse fibroblast (L929) cell culture. The results show that glucose concentration obtained from the SI system is comparable with that obtained from the conventional colorimetric method. This work can be further developed and applied for in vitro cell-based experiments in biomedical research.

Monitoring β-Fructofuranosidase Activity through Kluyveromyces marxianus in Bioreactor Using a Lab-Made Sequential Analysis System

“…In the context of this issue, automatic analyzers such as flow injection analysis systems (FIAs) have been proposed, which support routine determinations that demand frequent sampling [18]; another solution is the use of sequential injection analysis systems (SIAs), which allow discontinuous or intermittent modes of operation reducing reagents and sample consumption [19]. DNS methodologies have been applied in diverse flow systems to detect glucose content in soft drinks and wines [20] to detect L-lactate during lactic acid fermentation [21], for monitoring and controlling a lactose production process [22], and for cell culture applications [23]. SIA systems have been applied in bioprocesses to evaluate enzyme activity such as proteases [24], phospholipases [25], horseradish peroxidase [26,27], glucose oxidases [28], and lipases and esterase [29], among other applications [30,31].…”