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KMID : 0923620140140030164
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Immune Network
2014 Volume.14 No. 3 p.164 ~ p.170
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Characterization of Two Novel mAbs Recognizing Different Epitopes on CD43
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Kim So-Seul
Hong Jeong-won
Cho Woon-Dong
Moon Yoo-Ri
Yoon Sang-Soon
Kim Min-Young
Hong Kwon-Pyo
Lee Yong-Moon
Yi Jae-Hyuk
Ham Young-Jun
Rah Hyung-Chul
Kim Seung-Ryul
Song Hyung-Geun
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Abstract
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JL1, a specific epitope on CD43, is a potential biomarker for the diagnosis of acute leukemia. Although qualitative assays for detecting leukemia-specific CD43 exist, there is a need to develop quantitative assays for the same. Here, we devel-oped two novel monoclonal antibodies (mAbs), 2C8 and 8E10, recognizing different epitopes on CD43. These clones are capable of pairing with YG5, another mAb against JL1 epitope, because they were selectively obtained using sand-wich ELISA. Antigens recognized by 2C8 and 8E10 were confirmed as CD43 by western blotting using the CD43-hFC recombinant protein. When expression on various leukemic cell lines was investigated, 2C8 and 8E10 displayed a dis-parity in the distribution of the epitope. Enzyme assays re-vealed that these mAbs recognized a sialic acid-dependent epitope on CD43. Using normal thymus and lymph node par-affin-embedded tissues, we confirmed a difference in the epitopes recognized by the two mAbs that was predicted based on the maturity of the cells in the tissue. In summary, we developed and characterized two mAbs, 2C8 and 8E10, which can be used with YG5 in a sandwich ELISA for detect-ing leukemia-specific CD43.
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KEYWORD
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Leukemia, CD43, Epitope, Enzyme-Linked Immu-nosorbent Assay, Diagnosis
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