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大鼠-小鼠杂交瘤细胞在免疫抑制仓鼠体内的生长。一种从异种杂交瘤细胞系制备单克隆抗体的简便有效方法。

Growth of rat-mouse hybridoma cells in immunosuppressed hamsters. An easy and effective method to prepare monoclonal antibodies from heterohybridoma cell lines.

作者信息

Kayano T, Motoda R, Usui M, Ando S, Matuhasi T, Kurimoto M

机构信息

Hayashibara Biochemical Laboratories, Inc., Okayama, Japan.

出版信息

J Immunol Methods. 1990 Jun 12;130(1):25-31. doi: 10.1016/0022-1759(90)90295-7.

Abstract

Rat-mouse hybridoma cells producing anti-mouse IgE antibodies were intraperitoneally or subcutaneously inoculated into newborn or suckling hamsters receiving rabbit anti-hamster thymocyte globulin from the day of birth twice a week for at least 3 weeks. The hybridoma cells were found to grow in the abdominal cavity of the hamsters as ascites tumor or in subcutaneous tissue as solid tumor without loss of antibody-secreting activities. For the production of ascites, 2-week-old hamsters were preferable to newborn hamsters. In 3-week-old hamsters, the hybridoma cells could scarcely survive. The antibody titers of the ascites were determined to be 10(5)-10(6) in the ELISA and in the ability to neutralize the skin-sensitizing capacity of mouse IgE antibodies. The rat monoclonal antibodies were easily separated from ascites, serum or cell culture supernatant with affinity chromatography using Affigel protein A-Sepharose and anti-hamster IgG-Sepharose columns. The described method could be efficiently applicable for the proliferation of other hybridomas, such as human-human, human-mouse or hamster-mouse, etc.

摘要

将产生抗小鼠IgE抗体的大鼠 - 小鼠杂交瘤细胞腹腔内或皮下接种到新生或哺乳的仓鼠中,从出生当天起每周两次给予兔抗仓鼠胸腺细胞球蛋白,持续至少3周。发现杂交瘤细胞在仓鼠腹腔内以腹水瘤形式生长,或在皮下组织中以实体瘤形式生长,且不丧失抗体分泌活性。对于腹水的产生,2周龄的仓鼠比新生仓鼠更合适。在3周龄的仓鼠中,杂交瘤细胞几乎无法存活。腹水的抗体效价在ELISA中测定为10(5)-10(6),并且具有中和小鼠IgE抗体皮肤致敏能力。使用Affigel蛋白A - 琼脂糖和抗仓鼠IgG - 琼脂糖柱通过亲和色谱法可轻松从腹水、血清或细胞培养上清液中分离大鼠单克隆抗体。所描述的方法可有效应用于其他杂交瘤的增殖,如人 - 人、人 - 鼠或仓鼠 - 鼠等杂交瘤。

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