Development of a novel anti-human aspartyl-(asparaginyl) β-hydroxylase monoclonal antibody with diagnostic and therapeutic potential

Huyan,Ting; Li,Qi; Dong,Dan‑Dan; Yang,Hui; Xue,Xiao‑Ping; Huang,Qing‑Sheng

doi:10.3892/ol.2017.5642

Development of a novel anti-human aspartyl-(asparaginyl) β-hydroxylase monoclonal antibody with diagnostic and therapeutic potential

Authors:
- Ting Huyan
- Qi Li
- Dan‑Dan Dong
- Hui Yang
- Xiao‑Ping Xue
- Qing‑Sheng Huang
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Affiliations: Key Laboratory for Space Bioscience and Space Biotechnology, School of Life Sciences, Northwestern Polytechnical University, Xi'an, Shaanxi 710072, P.R. China
Published online on: January 25, 2017 https://doi.org/10.3892/ol.2017.5642
Pages: 1539-1546

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Abstract

Human aspartyl-(asparaginyl)-β-hydroxylase (HAAH) has recently been the subject of several studies, as it was previously observed to be overexpressed in numerous types of carcinoma cells and tissues in patient tumor samples. HAAH has been implicated in tumor invasion and metastasis, indicating that it may be an important target and biomarker for tumor diagnosis and treatment. However, the immunological tools currently available for the study of this protein, including monoclonal antibodies, are limited, as is the present knowledge regarding the role of HAAH in tumor therapy and diagnosis. In the present study, a recombinant C‑terminal domain of HAAH was expressed in Pichia pastoris and a novel monoclonal antibody (mAb) targeting HAAH (HAAH‑C) was constructed. Immunofluorescence and antibody‑dependent cellular cytotoxicity (ADCC) assays were used to demonstrate the specificity and ADCC activity of this antibody. The results demonstrated that this anti‑C‑terminal HAAH mAB, in combination with an existing anti‑N terminal HAAH mAb, exhibited a high response to native HAAH from carcinoma cell culture supernatant, as measured with a double antibody sandwich enzyme‑linked immunosorbent assay. This validated novel mAB‑HAAH‑C may prompt further studies into the underlying mechanisms of HAAH, and the exploration of its potential in tumor diagnosis and therapy.

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