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Receptor-based bioassay for functional IL-2 quantification.

Chuanwei Lu1, Shuangshuang Xu1, Xuelian Shan1

  • 1Analytical Science, WuXi Biologics, Hangzhou, China.

Journal of Pharmaceutical and Biomedical Analysis
|April 29, 2025
PubMed
Summary
This summary is machine-generated.

Manufacturing bioactive recombinant human interleukin-2 (rhIL-2) from inclusion bodies is challenging. A novel receptor-based ELISA accurately measures functional rhIL-2, aiding production and quality control.

Keywords:
Assay validationIL-2IL-2 receptor alphaIL-2 receptor beta & gammaInclusion bodyRefolding

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Area of Science:

  • Biotechnology
  • Immunology
  • Protein Chemistry

Background:

  • Interleukin-2 (IL-2) is crucial for immune cell proliferation and activation, underpinning immunotherapies for cancer and AIDS.
  • IL-2 is essential for in vitro immune cell culture, a key step in CAR-T and CAR-NK cell therapies.
  • Recombinant IL-2 production in E. coli offers cost and scalability advantages but often yields inactive inclusion bodies.

Purpose of the Study:

  • To address the lack of methods for assessing the biological activity of refolded recombinant IL-2 (rhIL-2).
  • To develop and present a novel receptor-based sandwich ELISA for quantifying functional rhIL-2.
  • To provide guidance for optimizing the manufacturing of bioactive rhIL-2 from prokaryotic expression systems.

Main Methods:

  • Development of a receptor-based sandwich ELISA assay.
  • Application of the ELISA to evaluate refolding efficiency of rhIL-2 produced from inclusion bodies.
  • Comparison of the receptor-based ELISA with traditional antibody-based methods.

Main Results:

  • The developed ELISA method accurately measures functional rhIL-2, reflecting its biological activity.
  • This assay provides a crucial tool for monitoring and optimizing the refolding process of rhIL-2.
  • The receptor-based approach is potentially adaptable for detecting modified rhIL-2 variants, such as PEGylated forms.

Conclusions:

  • A novel receptor-based ELISA effectively quantifies bioactive rhIL-2, overcoming limitations of existing methods.
  • This assay is vital for quality control in the bulk manufacturing of recombinant IL-2 using prokaryotic systems.
  • The method offers a robust platform for assessing functional protein refolding and can be extended to detect modified IL-2 variants.