Rapid Screen of IL-5/IL-5Rα Blocking Antibodies in the HEK293-IL-5Rα-CSF2RB Transfected Cell Line

• Published in: Biotechnology and bioprocess engineering Vol. 28; no. 4; pp. 612 - 622
• Main Authors: Li, Shijie, Han, Fei, Liu, Chang, Dai, Weiyan, Ke, Wenfeng, Chen, Yongqi, Fordjour, Eric, Yang, Yankun, Bai, Zhonghu
• Format: Journal Article
• Language: English
• Published: Seoul The Korean Society for Biotechnology and Bioengineering 01.08.2023; Springer Nature B.V; 한국생물공학회
• Subjects: Antibodies; Assaying; Asthma; Binding; Biotechnology; Blocking; Blocking antibodies; Chemistry; Chemistry and Materials Science; cytokine receptors; Cytokines; Drug development; Drugs; Eosinophils; Flow cytometry; fluorescence; humans; hybridomas; Industrial and Production Engineering; Interleukin 5; Interleukins; kidneys; Leukocytes (eosinophilic); Nanobodies; Receptors; Research Paper; Signal to noise ratio; Surface plasmon resonance; 생물공학; blocking antibody; asthma; interleukin-5/interleukin-5 receptor subunit alpha; cytokine receptor common subunit beta; fluorescence activated cell sorting
• ISSN: 1226-8372; 1976-3816
• DOI: 10.1007/s12257-022-0315-2

Interleukin-5 (IL-5) binding to interleukin-5 receptor subunit alpha (IL-5Rα) increases the number of eosinophils and enhances eosinophil activity. This leads to eosinophil tissue infiltration and damage to the lungs, ultimately resulting in exacerbation of asthma. Antibodies that block IL-5 binding to IL-5Rα are thought to play an important role in advanced asthma. Currently, key methods used to screen for targeted drugs are Surface Plasmon Resonance which is costly and anti-proliferation assays which are tedious and have a low signal-to-noise ratio. Here we describe a Fluorescence Activated Cell Sorting (FACS) assay, based on human embryonic kidney (HEK)-293 cells with stable expression of IL-5Rα and the cytokine receptor common subunit beta (CSF2RB). Cells co-expressing IL-5Rα and CSF2RB had a 16% increase in the ability to bind IL-5 compared to cells expressing only IL-5Rα. The optimal concentration of IL-5 for the FACS assay was 0.1 µg/mL. The established FACS was used to screen anti IL-5 nanobodies and hybridoma supernatants for candidate antibodies that block the IL-5/IL-5α interaction. When compared to anti-proliferation assays, this method saved up to 90% of the assay time, offering the advantage of rapidity and accuracy in vitro . The assay described here provides a novel approach for rapid screening of IL-5/IL-5Rα blocking antibodies in vitro to accelerate the development of drugs for asthma.