A novel artificially humanized anti‐cripto‐1 antibody suppressing cancer cell growth

Hiroko Ishii, Maram H. Zahra, Atushi Takayanagi, Masaharu Seno

Research output: Contribution to journalArticlepeer-review

6 Citations (Scopus)

Abstract

Cripto‐1 is a member of the EGF‐CFC/FRL1/Cryptic family and is involved in embryonic development and carcinogenesis. We designed a novel anti‐Cripto‐1 artificial antibody and assessed the recognition to the antigen and the potential to suppress the growth of cancer stem cells. First, single chain antibody clones were isolated by bio‐panning with the affinity to recombinant Cripto‐ 1 protein from our original phage‐display library. Then, the variable regions of heavy chain VH and light chain VL in each clone were fused to constant regions of heavy chain CH and light chain CL regions respectively. These fused genes were expressed in ExpiCHO‐S cells to produce artificial humanized antibodies against Cripto‐1. After evaluation of the expression levels, one clone was selected and the anti‐Cripto‐1 antibody was produced and purified. The purified antibody showed affinity to recombinant Cripto‐1 at 1.1 pmol and immunoreactivity to cancer tissues and cell lines. The antibody was available to detect the immunoreactivity in tissue microarrays of malignant tumors as well as in Cripto‐1 overexpressing cells. Simultaneously, the antibody exhibited the potential to suppress the growth of human colon cancer derived GEO cells overexpressing Cripto‐1 with IC50 at approximately 110 nM. The artificially humanized antibody is proposed to be a good candi-date to target cancer cells overexpressing Cripto‐1.

Original languageEnglish
Article number1709
Pages (from-to)1-14
Number of pages14
JournalInternational journal of molecular sciences
Volume22
Issue number4
DOIs
Publication statusPublished - Feb 2 2021

Keywords

  • Anti‐Cripto‐1 antibody
  • Artificial humanized antibody
  • Cell growth inhibition
  • Cripto‐1
  • Phage display library
  • Tissue‐micro array

ASJC Scopus subject areas

  • Catalysis
  • Molecular Biology
  • Spectroscopy
  • Computer Science Applications
  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Inorganic Chemistry

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