Development of novel-nanobody-based lateral-flow immunochromatographic strip test for rapid detection of recombinant human interferon α2b.

Journal: Journal of pharmaceutical analysis
Published Date: Jul 8, 2021

Abstract

Recombinant human interferon α2b (rhIFNα2b) is widely used as an antiviral therapy agent for the treatment of hepatitis B and hepatitis C. The current identification test for rhIFNα2b is complex. In this study, an anti-rhIFNα2b nanobody was discovered and used for the development of a rapid lateral flow strip for the identification of rhIFNα2b. RhIFNα2b was used to immunize an alpaca, which established a phage nanobody library. After five steps of enrichment, the nanobody I22, which specifically bound rhIFNα2b, was isolated and inserted into the prokaryotic expression vector pET28a. After subsequent purification, the physicochemical properties of the nanobody were determined. A semiquantitative detection and rapid identification assay of rhIFNα2b was developed using this novel nanobody. To develop a rapid test, the nanobody I22 was coupled with a colloidal gold to produce lateral-flow test strips. The developed rhIFNα2b detection assay had a limit of detection of 1 μg/mL. The isolation of I22 and successful construction of a lateral-flow immunochromatographic test strip demonstrated the feasibility of performing ligand-binding assays on a lateral-flow test strip using recombinant protein products. The principle of this novel assay is generally applicable for the rapid testing of other commercial products, with a great potential for routine use in detecting counterfeit recombinant protein products.

Authors

  • Xi Qin
    National Institutes for Food and Drug Control, Beijing, 100050, China.
  • Maoqin Duan
    National Institutes for Food and Drug Control, Beijing, 100050, China.
  • Dening Pei
    National Institutes for Food and Drug Control, Beijing, 100050, China.
  • Jian Lin
    Synthetic and Functional Biomolecules Center, Peking University, Beijing, 100871, China.
  • Lan Wang
    The Center of Psychosomatic Medicine, Sichuan Provincial Center for Mental Health, Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, Chengdu 611731, China.
  • Peng Zhou
    School of International Studies, Zhejiang University, Hangzhou, China.
  • Wenrong Yao
    National Institutes for Food and Drug Control, Beijing, 100050, China.
  • Ying Guo
    National Institutes for Food and Drug Control, Beijing, 100050, China.
  • Xiang Li
    Department of Radiology, Massachusetts General Hospital and Harvard Medical School, Boston, MA, United States.
  • Lei Tao
    National Institutes for Food and Drug Control, Beijing, 100050, China.
  • Youxue Ding
    National Institutes for Food and Drug Control, Beijing, 100050, China.
  • Lan Liu
    School of Statistics, University of Minnesota at Twin Cities.
  • Yong Zhou
    National Institutes for Food and Drug Control, Beijing, 100050, China.
  • Chuncui Jia
    National Institutes for Food and Drug Control, Beijing, 100050, China.
  • Chunming Rao
    National Institutes for Food and Drug Control, Beijing, 100050, China.
  • Junzhi Wang
    National Institutes for Food and Drug Control, Beijing, 100050, China.

Keywords

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