Novel anti-Tn single-chain Fv-Fc fusion proteins derived from immunized phage library and antibody Fc domain

Anticancer Res. 2010 Sep;30(9):3397-405.

Abstract

Tn[GalNAc(α1-3)-Ser/Thr] antigen, a tumor-associated carbohydrate antigen, is highly expressed in various tumors and an attractive candidate for cancer immunotherapy. The generation of an anti-Tn antibody is a first step toward the construction of new anticancer molecules. However, because of the simple and small conformation of the Tn molecule, it is difficult to generate an anti-Tn antibody for therapeutic use by conventional hybridoma technology. The purpose of this study was to isolate anti-Tn single-chain antibody fragments (scFv) by phage display technology from a novel immunised library, to attach an antibody constant region (Fc) and to convert them to scFv-Fc fusion proteins. The scFv-Fcs obtained here showed strict specificity against the Tn antigen and also showed antibody-dependent cellular cytotoxicity. These results suggest a potential use of this antibody generating method by phage display and indicate the potential of Fc-fusion proteins as therapeutic candidates.

MeSH terms

  • Animals
  • Antibody-Dependent Cell Cytotoxicity
  • Antigens, Tumor-Associated, Carbohydrate / immunology*
  • Cell Separation
  • Female
  • Flow Cytometry
  • Humans
  • Immunization
  • Immunoglobulin Fc Fragments / immunology*
  • Jurkat Cells
  • Mice
  • Mice, Inbred BALB C
  • Peptide Library*
  • Recombinant Fusion Proteins / chemical synthesis*
  • Recombinant Fusion Proteins / immunology
  • Reverse Transcriptase Polymerase Chain Reaction
  • Single-Chain Antibodies / immunology*

Substances

  • Antigens, Tumor-Associated, Carbohydrate
  • Immunoglobulin Fc Fragments
  • Peptide Library
  • Recombinant Fusion Proteins
  • Single-Chain Antibodies
  • Tn antigen