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Related Experiment Videos

Phage-derived monoclonal anti-Lu.

Martine Richard1, Josée Perreault, Pierre Gane

  • 1From Research and Development, Héma-Québec, Sainte-Foy, Québec, Canada.

Transfusion
|June 1, 2006
PubMed
Summary
This summary is machine-generated.

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Phage display technology successfully isolated a single-chain antibody fragment (scFv) recognizing the Lua blood group. This scFv was then reconstructed into a complete human monoclonal antibody (MoAb) with equivalent reactivity, marking the first MoAb specific for Lua.

Area of Science:

  • Immunology
  • Biotechnology
  • Hematology

Background:

  • Monoclonal antibodies (MoAbs) are increasingly used for blood typing, but some specificities are difficult to generate using hybridoma technology.
  • Phage display offers an alternative for isolating antibody fragments, potentially reducing analysis costs and improving phenotyping.

Purpose of the Study:

  • To isolate novel antibody fragments recognizing human red blood cell (RBC) specificities using phage display.
  • To reconstruct a functional human monoclonal antibody (MoAb) from an isolated fragment for blood group typing.

Main Methods:

  • A human single-chain variable fragment (scFv) phage display library was screened against red blood cells (RBCs) through three rounds of biopanning.
  • Selected scFv clones were analyzed, and one with Lua specificity was identified and reconstructed into a human IgG1 antibody.

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Main Results:

  • Three distinct clones were identified from the phage display library.
  • One clone demonstrated specific reactivity to the Lua blood group antigen.
  • The reconstructed human IgG1 MoAb showed indistinguishable reactivity to Lua compared to the parent scFv, confirmed by serologic assays and flow cytometry.

Conclusions:

  • Phage display technology enabled the successful isolation of an scFv with Lua specificity.
  • A complete human IgG1 MoAb with equivalent reactivity was reconstructed from the scFv.
  • This represents the first reported monoclonal antibody specific for the Lua blood group antigen.