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单克隆太阳麝香猫抗体:产生用于制备杂交瘤的融合伙伴

Monoclonal Suncus Antibodies: Generation of Fusion Partners to Produce - Hybridomas.

作者信息

Sado Yoshikazu, Inoue Satoko, Tomono Yasuko, Matsuyama Makoto, Fukushima Masaki, Oohashi Toshitaka, Jogahara Takamichi, Oda Sen-Ichi

机构信息

Shigei Medical Research Institute, 2117 Yamada, Minami-Ku, Okayama 701-0202, Japan.

Department of Molecular Biology and Biochemistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Kita-ku, Okayama 700-8558, Japan.

出版信息

Acta Histochem Cytochem. 2017 Apr 27;50(2):71-84. doi: 10.1267/ahc.17007. Epub 2017 Apr 15.

DOI:10.1267/ahc.17007
PMID:28522882
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5433937/
Abstract

We used suncus (; house musk shrew) to generate partner cells for cell fusion to produce suncus monoclonal antibodies. Suncus are insectivores that are genetically distant to rodents, and recognize antigens and epitopes that are not immunogenic in mice and rats, which are the animals most commonly used in basic life science research and from which monoclonal antibodies are usually produced. To date, monoclonal antibodies from suncus have not been generated due to the lack of a plasmacytoma fusion partner. To obtain suncus plasmacytoma cell lines suitable as a cell fusion partner, we injected suncus at both sides of the tail base with antigen emulsion, collected the lymph nodes and spleens, and cultured the cells to obtain immortalized lymphoid cell lines visually resembling mouse SP2/0-Ag14 myeloma cells. Three suncus immunized with the antigen provided 4 cell lines of suncus plasmacytoma, but they did not secrete immunoglobulins. Antibody-producing hybrid cells were generated from these cell lines using a cell fusion technique. Using one of the cell lines as a fusion partner, we obtained six lines of immunoglobulin-producing hybrid cells which secreted an unidentified monoclonal IgG. When these 6 lines were used as new fusion partners, we obtained several hybrid cell lines which secreted immunogen-specific monoclonal antibodies. These hybrid cells can be cloned and cryopreserved. We also obtained another good fusion partner which initially secreted antibody but later stopped doing so. These suncus-suncus hybrid cell lines will be useful for the production of suncus monoclonal antibodies.

摘要

我们使用臭鼩(家麝鼩)来生成用于细胞融合的伙伴细胞,以制备臭鼩单克隆抗体。臭鼩是食虫动物,在基因上与啮齿动物有很大差异,能识别在小鼠和大鼠中不具有免疫原性的抗原和表位,而小鼠和大鼠是基础生命科学研究中最常用的动物,也是通常用于生产单克隆抗体的动物。迄今为止,由于缺乏浆细胞瘤融合伙伴,尚未产生来自臭鼩的单克隆抗体。为了获得适合作为细胞融合伙伴的臭鼩浆细胞瘤细胞系,我们在臭鼩尾基部两侧注射抗原乳剂,收集淋巴结和脾脏,并培养细胞以获得在视觉上类似于小鼠SP2/0-Ag14骨髓瘤细胞的永生化淋巴细胞系。用该抗原免疫的三只臭鼩提供了4个臭鼩浆细胞瘤细胞系,但它们不分泌免疫球蛋白。使用细胞融合技术从这些细胞系中产生了产生抗体的杂交细胞。以其中一个细胞系作为融合伙伴,我们获得了6个分泌未鉴定单克隆IgG的产生免疫球蛋白的杂交细胞系。当将这6个细胞系用作新的融合伙伴时,我们获得了几个分泌免疫原特异性单克隆抗体的杂交细胞系。这些杂交细胞可以克隆和冷冻保存。我们还获得了另一个良好的融合伙伴,它最初分泌抗体,但后来停止分泌。这些臭鼩-臭鼩杂交细胞系将有助于臭鼩单克隆抗体的生产。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6c6/5433937/fa1496f0080d/AHC17007f10.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6c6/5433937/8b2bc05230d4/AHC17007f01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6c6/5433937/37272ef66881/AHC17007f02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6c6/5433937/59ce4dc169b5/AHC17007f03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6c6/5433937/8680f1b69c9f/AHC17007f04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6c6/5433937/0a71e8dcbe30/AHC17007f05.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6c6/5433937/27f6fabc7347/AHC17007f06.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6c6/5433937/219028179e74/AHC17007f07.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6c6/5433937/fb4253f602d7/AHC17007f08.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6c6/5433937/ba63f6683ca2/AHC17007f09.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6c6/5433937/fa1496f0080d/AHC17007f10.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6c6/5433937/8b2bc05230d4/AHC17007f01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6c6/5433937/37272ef66881/AHC17007f02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6c6/5433937/59ce4dc169b5/AHC17007f03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6c6/5433937/8680f1b69c9f/AHC17007f04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6c6/5433937/0a71e8dcbe30/AHC17007f05.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6c6/5433937/27f6fabc7347/AHC17007f06.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6c6/5433937/219028179e74/AHC17007f07.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6c6/5433937/fb4253f602d7/AHC17007f08.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6c6/5433937/ba63f6683ca2/AHC17007f09.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6c6/5433937/fa1496f0080d/AHC17007f10.jpg

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