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A protein chip approach for high‐throughput antigen identification and characterization
Author(s) -
Hu Shaohui,
Li Yu,
Liu Guozhen,
Song Qifeng,
Wang Li,
Han Yuning,
Zhang Yang,
Song Yali,
Yao Xiying,
Tao Yong,
Zeng Haipan,
Yang Huanming,
Wang Jian,
Zhu Heng,
Chen ZhiNan,
Wu Lin
Publication year - 2007
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200600923
Subject(s) - protein microarray , monoclonal antibody , antigen , proteomics , dna microarray , microarray , biology , microbiology and biotechnology , hepatocellular carcinoma , tissue microarray , computational biology , antibody , immunohistochemistry , gene , biochemistry , gene expression , cancer research , immunology
Proteomics research in humans and other eukaryotes demands a large number of high‐quality mAbs. Here, we report a new approach to produce high‐quality mAbs against human liver proteins using a combined force of high‐throughput mAb production and protein microarrays. After immunizing mice with live cells from human livers, we isolated 54 hybridomas with binding activities to human cells and identified the corresponding antigens for five mAbs via screening on a protein microarray of 1058 unique human liver proteins. Finally, we demonstrated that using the five mAbs we could characterize the expression profiles of their corresponding antigens by using tissue microarrays. Among them, we discovered that eIF1A expressed only in normal liver tissues, not in hepatocellular carcinoma in humans.