Surface-decorated S. cerevisiae for flow cytometric array immunoassay

Abstract

Our laboratory had developed a cell-based bio-bead for protein quantification. However, the selection of antibody in the above immunoassay is limited. This study describes a surface-decorated Saccharomyces cerevisiae for flow cytometric array immunoassay. S. cerevisiae was labeled with fluorescein isothiocyanate (FITC) and oxidized by sodium periodate, in which the saccharide group on the cytoderm outer layer was converted to an aldehyde group. In succession, adipic dihydrazide was bio-conjugated to the aldehyde group and glutaraldehyde bound to the hydrazide group. Phycoerythrin (PE)-labeled goat anti-mouse polyclonal antibody was used to assess the conjugation of mouse anti-human monoclonal antibody to surface-decorated S. cerevisiae. Cytokeratin 19 fragment (Cyfra21-1) and neuron-specific enolase (NSE) antigens were also employed to evaluate the flow cytometric array immunoassay based on surface-decorated S. cerevisiae. Flow cytometry demonstrated that FITC-barcoded S. cerevisiae as two legible populations. PE-labeled polyclonal antibody validated the coating of surface-decorated S. cerevisiae with the monoclonal antibody. The flow cytometric array immunoassays for Cyfra21-1 and NSE documented that the limit of detection (LOD) was at least 0.4 ng/mL. Precision and accuracy assessments appeared that the relative standard deviation (R.S.D.) was <15%, and the relative error (R.E.) ranged from 0.9 to 1.1. The correlation coefficient between this immunoassay and electrochemiluminescence immunoassay was 0.9622 for serum Cyfra21-1 and 0.9918 for serum NSE. In conclusion, the surface-decorated S. cerevisiae may be of use in flow cytometric array immunoassay.

Overcoming Aggregation-Caused Quenching by an Improved Porphyrin Hybrid and Its Application in Enhanced Electrochemiluminescence Biosensing

“…Therefore, it is crucial to be able to detect and diagnose NSCLC at an early stage. Cytokeratin-19 fragment (CYFRA21-1) is regarded as a rather effective bioindicator for the diagnosis and treatment of NSCLC, as it presents high levels in the serum of NSCLC patients. , Existing methods for serum CYFRA21-1 detection include enzyme-linked immunosorbent assay (ELISA), flow cytometry (FCM), chemiluminescence (CL), and electrochemical (EC). , It is worth mentioning that, compared to other methods, electrochemiluminescence (ECL) technology not only overcomes the drawbacks of time-consuming, cumbersome, and expensive steps but also combines the advantages of easy control, rapidity, simplicity, low cost, and high sensitivity of EC and CL, making it a great prospect for application. − Of these, the development of ECL emitters is crucial for ECL analysis performance.…”

Co-amplification of luminol-based electrochemiluminescence immunosensors based on multiple enzyme catalysis of bimetallic oxides CoCeOx and NiMnO3 for the detection of CYFRA21-1

“…Early detection and treatment of NSCLC are very crucial for cutting mortality and guaranteeing quality of life. At present, as a recognized biomarker for diagnosis of NSCLC, [2][3][4] cytokeratin 19 fragment antigen 21-1 (CYFRA21-1) has been analyzed by photoelectrochemical sensors, 5 electrochemical immunosensors, 6,7 fluorescent immunosensors, [8][9][10] flow cytometric array immunoassays, 11 and enzyme-linked immunosorbent assays. 12 However, these methods are often accompanied by disadvantages such as weak sensitivity, large background signals, and high costs.…”