Production of anti-recombinant human arrest defective 1 protein (hARD1) monoclonal antibodies for assaying human breast cancer tissues

Human arrest defective 1(hARD1) is an acetyltransferase catalyzing the N-terminal acetylation of proteins after translation. The high expression of hARD1 could be an indicator of the breast cancer. In current study, we produced an anti-hARD lp monoclonal antibody that could specifically recognize AR...

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Published inShengwu gongcheng xuebao Vol. 26; no. 1; p. 57
Main Authors Yu, Min, Wang, Zehua, Gong, Junli, Ma, Mingxing, Jiao, Yang, Huang, Weiwei, Lü, Qi, Li, Lin, Yang, Hui, Tan, Deyong
Format Journal Article
LanguageChinese
Published China 01.01.2010
Subjects
Acetyltransferases - genetics
Acetyltransferases - immunology
Animals
Antibodies, Monoclonal - biosynthesis
Antibodies, Monoclonal - genetics
Antibodies, Monoclonal - immunology
Biomarkers, Tumor - biosynthesis
Biomarkers, Tumor - immunology
Breast Neoplasms - immunology
Breast Neoplasms - metabolism
Breast Neoplasms - pathology
Escherichia coli - genetics
Escherichia coli - metabolism
Female
Humans
Immunization
Mice
Mice, Inbred BALB C
N-Terminal Acetyltransferase A
N-Terminal Acetyltransferase E
Recombinant Proteins - biosynthesis
Recombinant Proteins - genetics
Recombinant Proteins - immunology
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Summary:Human arrest defective 1(hARD1) is an acetyltransferase catalyzing the N-terminal acetylation of proteins after translation. The high expression of hARD1 could be an indicator of the breast cancer. In current study, we produced an anti-hARD lp monoclonal antibody that could specifically recognize ARD1 in breast cancer tissues by using the immunohistochemical assay. The full-length His-tag hARD1 protein (1-235 aa) was over-expressed in Escherichia coli, and purified recombinant protein was injected into Balb/c mice to perform immunization procedure. Eight stable positive monoclonal cell lines were isolated. ELISA results demonstrated that all light chains of antibodies were kappa, and the heavy chains displayed three subtypes IgG1, IgG2a and IgG2b, respectively. A monoclonal antibody, which could specifically recognize hARD1 protein in breast cancer tissues, was identified by screening different cancer tissues using antibody-specificity method. Further, the specificity of the antibody was confirmed by Western
ISSN:1000-3061