Screening high affinity monoclonal antibody producing hybridomas using a graphene oxide-based fluorescence biosensor

Abstract

Screening monoclonal antibody (mAb) producing hybridomas by ELISA using microplates is time-consuming and expensive. The combination of biosensing and flow cytometry represents a promising rapid, high-throughput method for screening mAb producing clones. Based on the assembly of graphene oxide (GO) and fluorescein isothiocyanate (FITC)-labeled peptide that included the N-terminal portion of selenoprotein K (SELENOK) antigen epitope, we designed a novel GO fluorescence biosensor for screening high affinity mAb producing hybridomas. The FITC-labeled peptide was strongly adsorbed on the surface of GO, which effectively quenched the FITC fluorescence. The specific SELENOK mAb competitively bound to FITC-Peptide, resulting in FITC-Peptide release from the GO surface and fluorescence recovery. This sensor was successfully utilized to sensitively detect SELENOK mAb with low detection limit of 0.25 U. In addition, we employed this FITC-Peptide/GO biosensor to in situ visualizing and high-throughput screening SELENOK mAb hybridomas (SMAHs) at the single-cell level. The proposed sensor is simple, relatively inexpensive, highly specific, and applicable for cell sort assay, demonstrating the effective use for peptide-antibody interactions in the specific screening for other mAb hybridomas libraries.