Bacterial expression and characterization of an anti-CD22 single-chain antibody fragment

Protein Expr Purif. 2020 Jun:170:105594. doi: 10.1016/j.pep.2020.105594. Epub 2020 Feb 5.

Abstract

Single-chain variable fragment (scFv) antibodies are fusion proteins of the variable regions of the heavy and light chains of immunoglobulins connected with a short linker peptide. They possess unique and superior features compared to whole antibodies for immunotherapy of various carcinomas, including hematologic B-cell malignancies. In the presented study we obtained efficient production of the recombinant anti-CD22 scFv in Escherichia coli expression system. The active recombinant protein was successfully recovered from inclusion bodies. Assays were performed to assess the in vitro targeting properties and specificity of the obtained anti-CD22 scFv antibody in the CD22 positive and negative lymphoma cell lines.

Keywords: Bacterial expression system; CD22; Recombinant protein production; Single-chain variable fragment.

MeSH terms

  • Cell Line, Tumor
  • Cloning, Molecular
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Fluorescein-5-isothiocyanate / chemistry
  • Fluorescent Dyes / chemistry
  • Gene Expression
  • Genetic Vectors / chemistry
  • Genetic Vectors / metabolism
  • Humans
  • Immunoconjugates / chemistry*
  • Immunoconjugates / metabolism
  • Inclusion Bodies / chemistry
  • K562 Cells
  • Lymphocytes / metabolism*
  • Lymphocytes / pathology
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Sialic Acid Binding Ig-like Lectin 2 / chemistry
  • Sialic Acid Binding Ig-like Lectin 2 / genetics*
  • Sialic Acid Binding Ig-like Lectin 2 / metabolism
  • Single-Chain Antibodies / chemistry
  • Single-Chain Antibodies / genetics*
  • Single-Chain Antibodies / metabolism

Substances

  • CD22 protein, human
  • Fluorescent Dyes
  • Immunoconjugates
  • Recombinant Proteins
  • Sialic Acid Binding Ig-like Lectin 2
  • Single-Chain Antibodies
  • Fluorescein-5-isothiocyanate