Auto-Lectin Dotcoding by Two Octopuses: Rapid Analysis of Fluorescence-Labeled Glycoproteins by an 8-channel Fully-Automatic Bead Array Scanner with a Rolling-Circle Detector.
Hiroko ShimazakiKazuhiro NakamuraAyaka OnoOsamu SegawaKazumi SawakamiMichinori KoizukaMakoto HirayamaKanji HoriHideji TajimaAtsushi KunoPublished in: Analytical chemistry (2023)
Protein glycosylation is a crucial factor that must be evaluated in biological pharmaceuticals. The glycoform profile of a protein can vary depending on the conditions of the cultivation, purification process, and the selection of a host cell. Lectin microarrays are reliable bioanalytical methods used in the early phases of bioprocesses for the detection of glycosylation. The concept of a fully automated glycan detection with a bead array has been previously reported; however, no simple system has been constructed on fluorescence-based detection using a microarray. Here, we present a fully automated detection system equipped with a novel fluorescence detector for a 13-lectin bead array with a single tip. The lattice-like arrangement of a set of fibers proximate to the tip of the light emitting diode and photomultiplier tube detector minimized the noise caused by the reflection of incident light on the plastic capillary tip and bead. A unique rolling-circle fiber unit with quadruple lattices stacked in two layers realizes the 8-parallel automeasurement with a drastic reduction in scanning time and machine size. The 8-glycan profiles obtained automatically within 25 min were identical with those obtained with the conventional lectin microarray after overnight incubation. The signals obtained were represented as lectin dotcodes. Therefore, autolectin dotcoding assisted by the twin 8 legs named as "detection and irradiation octopuses" may be a rapid glyco-evaluation system during the production and development of biopharmaceuticals.
Keyphrases
- loop mediated isothermal amplification
- high throughput
- deep learning
- label free
- real time pcr
- high resolution
- machine learning
- single molecule
- computed tomography
- sensitive detection
- stem cells
- cardiovascular disease
- single cell
- cell therapy
- mass spectrometry
- radiation therapy
- type diabetes
- wastewater treatment
- mesenchymal stem cells
- positron emission tomography
- small molecule
- radiation induced
- amino acid
- helicobacter pylori infection