Identification of a mimotope for circulating anti-cytokeratin 8/18 antibody and its usage for the diagnosis of breast cancer

Heo,Chang-Kyu; Hwang,Hai-Min; Ruem,Ah; Yu,Dae-Yeul; Lee,Ju   Yeon; Yoo,Jong  Shin; Kim,In Gyu; Yoo,Hyang  Sook; Oh,Sejeong; Ko,Jeong  Heon; Cho,Eun-Wie

doi:10.3892/ijo.2012.1679

Identification of a mimotope for circulating anti-cytokeratin 8/18 antibody and its usage for the diagnosis of breast cancer

Authors:
- Chang-Kyu Heo
- Hai-Min Hwang
- Ah Ruem
- Dae-Yeul Yu
- Ju Yeon Lee
- Jong Shin Yoo
- In Gyu Kim
- Hyang Sook Yoo
- Sejeong Oh
- Jeong Heon Ko
- Eun-Wie Cho
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Affiliations: Cancer Biomarkers Development Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Republic of Korea, Aging Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Republic of Korea, Division of Mass Spectrometry Research, Korea Basic Science Institute, Daejeon, Republic of Korea, Department of Radiation Biology, Environmental Radiation Research Group, Korea Atomic Energy Research Institute, Daejeon, Republic of Korea, Department of Surgery, College of Medicine, The Catholic University of Korea, Inchon, Republic of Korea
Published online on: October 26, 2012 https://doi.org/10.3892/ijo.2012.1679
Pages: 65-74
Copyright: © Heo et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

A novel circulating tumor-associated autoantibody, K94, obtained from a hepatocellular carcinoma (HCC) mouse model was characterized. The target antigen of K94 autoantibody was expressed in various tumor cell lines including liver cancer, and its secretion was detectable using MCF-7 breast carcinoma cells. Proteomic analysis revealed that the protein bands reactive to K94 included cytokeratin (CK) 8 and 18, which are known to be related to tumorigenesis and form a heterotypic complex with each other. However, K94 showed no activity toward CK8 or CK18 separately. The epitope of the K94 antibody was only presented by a complex between CK8 and CK18, which was confirmed by analysis using recombinant CK8 and CK18 proteins. To formulate an assay for anti-CK8/18 complex autoantibody, a mimotope peptide reactive to K94 was selected from loop-constrained heptapeptide (-CX7C-) display phage library, of which sequence was CISPDAHSC (K94p1). A mimotope enzyme-linked immunosorbent assay (ELISA) using phage-displayed K94p1 peptide as a coating antigen was able to discriminate breast cancer (n=30) patients from normal subjects (n=30) with a sensitivity of 50% and a specificity of 82.61%. CA15.3 was detected at very low levels in the same breast cancer subjects and did not discriminate breast cancer patients from normal subjects, although it is a conventional biomarker of breast cancer. These results suggest that a mimotope ELISA composed of K94p1 peptide may be useful for the diagnosis of breast cancer.

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