A Novel Artificially Humanized Anti-Cripto-1 Antibody Suppressing Cancer Cell Growth
Open Access
- 8 February 2021
- journal article
- research article
- Published by MDPI AG in International Journal of Molecular Sciences
- Vol. 22 (4), 1709
- https://doi.org/10.3390/ijms22041709
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 candidate to target cancer cells overexpressing Cripto-1.This publication has 36 references indexed in Scilit:
- CHO Quasispecies—Implications for Manufacturing ProcessesProcesses, 2013
- Phage-encoded combinatorial chemical libraries based on bicyclic peptidesNature Chemical Biology, 2009
- Isolation and Characterization of Phage-Displayed Single Chain Antibodies Recognizing Nonreducing Terminal Mannose Residues. 1. A New Strategy for Generation of Anti-Carbohydrate AntibodiesBiochemistry, 2007
- Cripto-1: a multifunctional modulator during embryogenesis and oncogenesisOncogene, 2005
- Amphiregulin anti‐sense oligodeoxynucleotides inhibit growth and transformation of a human colon carcinoma cell lineInternational Journal of Cancer, 1995
- Phage antibodies: filamentous phage displaying antibody variable domainsNature, 1990
- Filamentous Fusion Phage: Novel Expression Vectors That Display Cloned Antigens on the Virion SurfaceScience, 1985
- Establishment and characterization of a cell line of human breast carcinoma originEuropean Journal of Cancer (1965), 1979
- Characteristics of a Human Cell Line Transformed by DNA from Human Adenovirus Type 5Journal of General Virology, 1977
- Determination of the helix and β form of proteins in aqueous solution by circular dichroismBiochemistry, 1974