Microquantitation of Van Gogh-like Protein 1 by Using Antibody-Conjugated Magnetic Beads
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Original Article
Microquantitation of Van Gogh-like Protein 1 by Using Antibody-Conjugated Magnetic Beads Su-Jung Yeom1,†, Shin-Yeong Lee1,†, İsa Taş1,†, Mijin Lee1, So-Yeon Park1, Sang-Chul Jung2, Kyung Keun Kim3 & Hangun Kim 1,* Received: 18 December, 2018 / Accepted: 21 January, 2019 / Published online: 15 April, 2019 ⒸThe Korean BioChip Society and Springer 2018
Abstract Van Gogh-like protein 1 (VANGL1) is an integral membrane protein that has a decisive effect on invasion, migration, and metastasis of tumor cells in various cancers including colorectal cancer. Elevation of VANGL1 level in cancer tissue is often observed with progressive and metastatic colorectal cancer. Therefore, detection of VANGL1 level in patient specimens can be used as a diagnostic marker for colorectal cancer screening. In this study, we developed magnetic beads that conjugate the VANGL1 antibody, which can be used for quantitative analysis of VANGL1. The procedure for bead preparation was optimized, and detection analysis was validated by using Caco2 and HT29 colorectal cancer cell lysates. Through the procedure, VANGL1 level from cell lysates of Caco2 and HT29 cells were quantified to estimate whether the antibody-conjugated magnetic bead can be used to trace amounts of VANGL1. Results from VANGL1 level obtained by using the antibody-conjugated magnetic beads suggest that the procedure has high precision and sensitivity in analyzing VANGL1. In conclusion, the results indicate that the method is appropriate for microquantitation of VANGL1 from patient specimens.
1 College of Pharmacy, Sunchon National University, Sunchon 57922, Korea 2 Department of Environmental Engineering, Sunchon National University, Sunchon 57922, Korea 3 Department of Pharmacology, Chonnam National University Medical School, Hwasun 58128, Korea † These three authors contributed equally to this work. *Correspondence and requests for materials should be addressed to H. Kim ( [email protected])
Keywords: Colorectal cancer specific antigen, VANGL1, Protein, Quantitation, Diagnosis
Introduction Wnt signaling pathway has critical functions in tumorigenesis through the canonical Wnt/β-catenin and in invasion, metastasis, and angiogenesis through noncanonical Wnt/planar cell polarity (PCP) signaling1. One of the core component of Wnt/PCP, Van Gogh-like protein 1 (VANGL1), is located on human chromosome 1p13 and encodes a transmembrane protein that interacts with other PCP proteins Disheveled, Prickle, and Frizzled, a tumor metastasis suppressor KAI1 protein, and an intestinal epithelial restitution factor ITF protein2,3. Clinically, VANGL1 expression was significantly higher in colorectal cancer (CRC) tissues from the advanced stage (III, IV) than that of stage I and associated with lymphovascular invasion, depth of invasion, lymph node metastasis, and poor survival4,5. The expression level of VANGL1 in colorectal cancer tissues can be used for colorectal cancer screening, prognosis prediction and malignancy judgment of colorectal cancer5–8. The levels of VANG
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