Smartphone Cortex Controlled Real-Time Image Processing and Reprocessing for Concentration Independent LED Induced Fluorescence Detection in Capillary Electrophoresis

Abstract

We present the application of a smartphone anatomy based technology in the field of liquid phase bioseparations, particularly in capillary electrophoresis. A simple capillary electrophoresis system was built with LED induced fluorescence detection and a credit card sized minicomputer to prove the concept of real time fluorescent imaging (zone adjustable time-lapse fluorescence image processor) and separation controller. The system was evaluated by analyzing under- and overloaded aminopyrenetrisulfonate (APTS)-labeled oligosaccharide samples. The open source software based image processing tool allowed undistorted signal modulation (reprocessing) if the signal was inappropriate for the actual detection system settings (too low or too high). The novel smart detection tool for fluorescently labeled biomolecules greatly expands dynamic range and enables retrospective correction for injections with unsuitable signal levels without the necessity to repeat the analysis.

Applications of capillary electrophoresis for biopharmaceutical product characterization

“…Besides human IgG, the platform was also useful for sequencing Etanercept, a fusion protein‐based biopharmaceutical. Researchers have recently reported a method for image processing of the fluorescent signal in real‐time using LED‐induced fluorescence detectors (LedIF) [75]. The system allowed retrospective correction of sample injection for under‐ and over‐loaded labeled glycans.…”

“…Method developments in mAb glycosylation analysis using CE were reported using commercial human IgG as the model sample [74][75][76][77][78][79][80][81][82]. For example, in one study, the hardware components of the CE instrument itself were used to develop semi and fully automated CE-LIF-based methods for carbohydrate sequencing of released glycans [74].…”

Greening Capillary Electrophoresis, a promising sprout of Separation Science toward sustainability

“…• System A: Separations were performed with a previously described custom-made capillary electrophoresis unit, equipped with LED-induced fluorescence imaging detection system (Szarka and Guttman, 2017). Effective capillary length was 20 cm (total length 30 cm), 50 µm ID Bare Fused Silica capillary.…”

“…Two-stage manual electrokinetic sample injection method was applied to utilize stacking effect: (i) 1 kV for 1 s water injection, (ii) sample injection at 3 kV for 3 s. The N-CHO separation gel buffer (SCIEX) served as background electrolyte. A custom written software package was used for data acquisition and processing, following our method described earlier (Szarka and Guttman, 2017). • System B: Reference separations were performed on a PA800 Plus Pharmaceutical Analysis System (SCIEX).…”

“…In this paper we intend to further emphasize the sustainability of capillary electrophoresis with LED-induced fluorescence detection. For this, we compare the same PNGase F released, APTS-labeled immunoglobulin G N-glycan separation results obtained with System A, a home built low consumption CE device, operated semi automatically (Szarka and Guttman, 2017) and System B, a fully automated, high end commercial device. Our analysis shows that with some limitations both techniques show potential in the separation milieu.…”