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Development of an anti-CD45RA-quantum dots conjugated scFv to detect leukemic cancer stem cells

  • Shima Moradi-Kalbolandi
  • Fariba Dashtestani
  • Malihe Salehi
  • Neda Jalili
  • Keivan Majidzadeh-A
  • Reza Rahighi
  • Amir Yadegari
  • Leila FarahmandEmail author
Original Article
  • 77 Downloads

Abstract

Leukemic cancer stem cells (LSCs), aberrantly overexpressing CD45RA are among the major causes of relapse following chemotherapy in patients with acute myeloid leukemia and serve as a highly sensitive marker for predicting relapse occurrence following chemotherapy. The main purpose of current study was to develop a sensitive approach for detecting LSCs based on a conjugate of an anti-CD45 scFv and quantum dot. The variable light and heavy chain sequences of a recently developed anti-CD45RA monoclonal antibody were derived from hybridoma cells and PCR amplified to construct scFv. Following insertion of scFv gene into a pET32a-lic vector and expression in Escherichia coli and purification, the purified scFv, was conjugated with carbon dots (C dots) and used for the detection of CD45RA +cells while CD45RA-cells served as negative control. Subsequently, Functional activity of the conjugate was analyzed by flow cytometry and ICC to detect the cell surface antigen binding and detection ability. Based on results, purified CD45RA scFv conjugated C dots could specifically recognize CD45RA positive cells, but not any CD45RA negative ones. In conclusion, here we developed a low-cost but very efficient approach for detection of CD45RA positive cells including LSCs.

Keywords

AML CD45RA scFv Leukemic cancer stem cell Carbon dots Conjugation 

Notes

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Shima Moradi-Kalbolandi
    • 1
  • Fariba Dashtestani
    • 1
  • Malihe Salehi
    • 1
  • Neda Jalili
    • 1
  • Keivan Majidzadeh-A
    • 1
  • Reza Rahighi
    • 2
  • Amir Yadegari
    • 3
  • Leila Farahmand
    • 1
    Email author
  1. 1.Recombinant Proteins Department, Breast Cancer Research Center, Motamed Cancer InstituteACECRTehranIran
  2. 2.Department of Research and DevelopmentSharif Ultra High Nanotechnologists (SUN) CompanyTehranIran
  3. 3.Department of Developed SciencesMarquette UniversityMilwaukeeUSA

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