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Development and characterization of a camelid single-domain antibody directed to human CD22 biomarker

Fatemeh Faraji1,2, Nader Tajik1,2, Mahdi Behdani3

  • 1Department of Immunology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran.

Biotechnology and Applied Biochemistry
|March 16, 2018
PubMed

Insights

Researchers developed a novel anti-CD22 variable domain of camelid single-chain antibody (VHH) for B-cell malignancies. This highly specific and high-affinity VHH shows promise for diagnosing and treating B-cell disorders.

Area of Science:

  • Immunology
  • Biochemistry
  • Biotechnology

Background:

  • CD22 is a B-cell-specific surface glycoprotein modulating B-cell function, survival, and apoptosis.
  • Targeting CD22 is a promising strategy for B-cell malignancies and disorders.
  • Camelid single-chain variable fragments (VHH, nanobodies) offer advantages like small size, stability, and high affinity.

Purpose of the Study:

  • To develop and characterize a novel VHH specific for the CD22 antigen.
  • To evaluate the binding affinity and specificity of the anti-CD22 VHH to CD22+ B cells.

Main Methods:

  • Screening of an immunized phage display library to identify anti-CD22 VHH.
  • Western blotting to confirm molecular size of the produced VHH.
  • Flow cytometry and cell lysate assays to evaluate binding affinity and specificity.

Main Results:

  • A novel anti-CD22 VHH was successfully developed and characterized.
  • The VHH exhibited a molecular size of approximately 17 kDa and a binding affinity of 9 x 10^-9 M.
  • The VHH demonstrated high specificity, recognizing natural CD22 antigen on cell surfaces (93%) and in cell lysates.

Conclusions:

  • The developed anti-CD22 VHH possesses high affinity and specificity for the CD22 antigen.
  • This VHH is a potential candidate for diagnostic and therapeutic applications in B-cell disorders and malignancies.

Keywords:
CD22VHHcamelid single-domain antibody

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