PMab-213: A Monoclonal Antibody for Immunohistochemical Analysis Against Pig Podoplanin
PMab-213: A Monoclonal Antibody for Immunohistochemical Analysis Against Pig Podoplanin
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DOI：
10.1089/mab.2018.0048
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发表时间：
2019-02-01
影响因子：
--
通讯作者：
Kaneko, Mika K.
中科院分区：
文献类型：
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作者：
Kato, Yukinari;Yamada, Shinji;Kaneko, Mika K.
Podoplanin（PDPN）已知在正常组织中表达，包括淋巴管内皮细胞、肾足细胞和I型肺泡细胞。已经建立了针对人、小鼠、大鼠、兔、狗、猫和牛PDPN的单克隆抗体（mAb）;然而，缺乏针对猪PDPN（pPDPN）的mAb。在本研究中，用过表达pPDPN的中国仓鼠卵巢（CHO）-K1细胞（CHO/pPDPN）免疫小鼠，并通过流式细胞术筛选产生针对pPDPN的mAb的杂交瘤。其中一株单抗PMab-213（IgG（2b），kappa）可通过流式细胞术特异性检测CHO/pPDPN细胞，通过western blot检测pPDPN。测定PMab-213对CHO/pPDPN的KD为2.1 x 10（-9）M，表明对CHO/pPDPN具有高亲和力。此外，PMab-213通过免疫组织化学强烈染色淋巴管内皮细胞、肾足细胞和I型肺泡细胞。PMab-213有望用于研究pPDPN的功能。
Podoplanin (PDPN) is known to be expressed in normal tissues, including lymphatic endothelial cells, renal podocytes, and type I lung alveolar cells. Monoclonal antibodies (mAbs) against human, mouse, rat, rabbit, dog, cat, and bovine PDPN have already been established; however, mAbs against pig PDPN (pPDPN) are lacking. In the present study, mice were immunized with pPDPN-overexpressing Chinese hamster ovary (CHO)-K1 cells (CHO/pPDPN), and hybridomas producing mAbs against pPDPN were identified by flow cytometric screening. One of the mAbs, PMab-213 (IgG(2b), kappa), could specifically detect CHO/pPDPN cells through flow cytometry and detect pPDPN through western blot analysis. K-D of PMab-213 for CHO/pPDPN was determined to be 2.1 x 10(-9) M, indicating a high affinity for CHO/pPDPN. Furthermore, PMab-213 strongly stained lymphatic endothelial cells, renal podocytes, and type I lung alveolar cells through immunohistochemistry. PMab-213 is expected to be useful in investigating the function of pPDPN.