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Updated: Feb 4, 2026

Single-cell Screening Method for the Selection and Recovery of Antibodies with Desired Specificities from Enriched Human Memory B Cell Populations
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Generation and characterization of avian-derived anti-human CD19 single chain fragment antibodies

Keng-Chang Tsai1,2, Chen-Wei Chiang3, Yan-Ni Lo3

  • 1National Research Institute of Chinese Medicine, Ministry of Health and Welfare , Taipei , Taiwan.

Animal Biotechnology
|September 29, 2018
PubMed

Insights

Researchers developed chicken-derived anti-CD19 single-chain variable fragments (scFv) for leukemia therapy. The S2 scFv clone specifically binds CD19, offering potential for targeted cancer treatments and antibody engineering.

Area of Science:

  • Immunology
  • Biotechnology
  • Molecular Biology

Background:

  • Cluster of differentiation 19 (CD19) is a key target in leukemia due to its high expression.
  • Chicken antibodies offer a valuable platform for therapeutic protein development.
  • Phage display technology is a powerful tool for antibody discovery.

Purpose of the Study:

  • To generate and characterize novel anti-CD19 single-chain variable fragments (scFv) from immunized chickens.
  • To investigate the binding affinity and specificity of the developed scFv against CD19.
  • To explore the structural properties of chicken antibody complementarity-determining regions (CDRs) for antibody engineering.

Main Methods:

  • Immunization of chickens and construction of a large-scale scFv phage display library (2.5 × 10^8 diversity).
  • Panning of the library to isolate CD19-specific scFv clones, including clone S2.
  • Characterization of scFv S2 binding using flow cytometry and surface plasmon resonance (SPR).
  • Analysis of complementarity-determining regions (CDRs) through hypermutation and molecular modeling.

Main Results:

  • Isolation and characterization of a specific anti-CD19 scFv clone, designated S2.
  • Verification of S2 binding to CD19 on ARH-77 leukemia cells via flow cytometry.
  • Determination of S2 binding affinity to CD19 as 6.9 × 10^-8 M by SPR.
  • Identification of antigen-driven hypermutation in S2's CDRs, suggesting an effective immune response.

Conclusions:

  • Chicken-derived scFv S2 demonstrates specific binding and affinity for the CD19 target.
  • The study provides insights into the structural characteristics of chicken antibody CDRs.
  • Findings contribute to the development of novel antibody-based therapeutics and engineering strategies for leukemia.
Keywords:
Human cluster of differentiation 19leukemiamolecular modelingphage display technologysingle chain variable fragment

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