Analysis of the clinical significance of DCLK1+ colorectal cancer using novel monoclonal antibodies against DCLK1

OncoTargets and Therapy
Tianqi DaiAishun Jin

Abstract

Doublecortin-like kinase 1 (DCLK1) is considered a putative tumor stem cell (TSC) marker and a promising therapeutic target, as DCLK1+ progeny cells exhibit high expression in tumors. However, the biological function of DCLK1+ cells in tumorigenesis and tumor progression remains unclear. We generated rabbit monoclonal antibodies (mAbs) against DCLK1, DCLK1-42, and DCLK1-87 mAbs, using a novel chip-based immunospot array assay on a chip system. First, the specificity of two mAbs to DCLK1 was confirmed by Western blot, which were bound to DCLK1-long in normal colon cells and to DCLK1-short in a cancer cell line as well as colorectal cancer (CRC) cells. Precise localization analysis using immunofluorescence revealed that both mAbs had cytoplasmic signal and exhibited a high degree of overlap with microtubules. Furthermore, bacterial display technology indicated that the antigenic epitope region of DCLK1-87 mAb was consistent with that of a commercial anti-DCLK1 polyclonal antibody. In addition, DCLK1-42 mAb has the common polyclonal antibody characteristic of binding to more than one site on DCLK1. By immunohistochemistry, it was found that DCLK1-87 mAb was more specific for DCLK1+ cell labeling than a commercial anti-DCLK1 polycl...Continue Reading

Citations

Jul 17, 2020·Journal of Cancer Research and Clinical Oncology·Arkadiusz GzilŁukasz Szylberg
Jun 28, 2019·Journal of Drug Targeting·Meng-Yan WangRong-Guang Shao
Sep 27, 2020·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Lorenzo NeviDomenico Alvaro

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Methods Mentioned

BETA
chip
electrophoresis
PCR
xenografts
fluorescence microscopy
flow cytometry
chips

Software Mentioned

GraphPad Prism
SPSS Statistics

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