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Yoon, Joo, Kim, Kang, Kim, Lee, Kwon, Kim, Choe, and Kim: Characterization of a Monoclonal Antibody Specific to Human Siah-1 Interacting Protein
Original Article

Immune Network 2004; 4(1): 23-30.

Published online: 31 March 2004

DOI: https://doi.org/10.4110/in.2004.4.1.23

Characterization of a Monoclonal Antibody Specific to Human Siah-1 Interacting Protein

Sun Young Yoon1, Joung Hyuck Joo1, Joo Heon Kim2, Ho Bum Kang1, Jin Sook Kim1, Younghee Lee1, Do Hwan Kwon1, Chang Nam Kim3, In Seong Choe1, Jae Wha Kim1

1Laboratory of Cell Biology, Korea Research Institute of Bioscience and Biotechnology (KRIBB), P.O. Box 115, Daejeon, Republic of Korea.

2Department of Pathology, Eulji University School of Medicine, Daejeon, Republic of Korea.

3Department of Surgery, Eulji University School of Medicine, Daejeon, Republic of Korea.

Corresponding Author (choerncbg@kribb.re.kr)

Copyright © 2004 The Korean Association of Immunologists

(open-access):

This is an open access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background

A human orthologue of mouse S100A6-binding protein (CacyBP), Siah-1-interacting protein (SIP) had been shown to be a component of novel ubiquitinylation pathway regulating β-catenin degradation. The role of the protein seems to be important in cell proliferation and cancer evolution but the expression pattern of SIP in actively dividing cancer tissues has not been known. For the elucidation of the role of SIP protein in carcinogenesis, it is essential to produce monoclonal antibodies specific to the protein.

Methods

cDNA sequence coding for ORF region of human SIP gene was amplified and cloned into an expression vector to produce His-tag fusion protein. Recombinant SIP protein and monoclonal antibody to the protein were produced. The N-terminal specificity of anti-SIP monoclonal antibody was conformed by immunoblot analysis and enzyme linked immunosorbent assay (ELISA). To study the relation between SIP and colon carcinogenesis, the presence of SIP protein in colon carcinoma tissues was visualized by immunostaining using the monoclonal antibody produced in this study.

Results

His-tag-SIP (NSIP) recombinant protein was produced and purified. A monoclonal antibody (Korea patent pending; #2003-45296) to the protein was produced and employed to analyze the expression pattern of SIP in colon carcinoma tissues.

Conclusion

The data suggested that anti-SIP monoclonal antibody produced here was valuable for the diagnosis of colon carcinoma and elucidation of the mechanism of colon carcinogenesis.

Keywords: Siah-1 interacting protein(SIP), S100A6, monoclonal antibody, colorectal cancer

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