Lymphocytes from enlarged iliac lymph nodes as fusion partners for the production of monoclonal antibodies after a single tail base immunization attempt.
Lymphocytes from enlarged iliac lymph nodes as fusion partners for the production of monoclonal antibodies after a single tail base immunization attempt.
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DOI：
10.1267/ahc.06001
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发表时间：
2006-07-01
影响因子：
2.4
通讯作者：
Omori H
中科院分区：
文献类型：
--
作者：
Sado Y;Inoue S;Tomono Y;Omori H
建立了一种新的制备鼠源性单克隆抗体杂交瘤的方法，称为“小鼠髂淋巴结法”。来自在尾基部肌内注射抗原和弗氏佐剂的乳液的小鼠的增大的髂淋巴结的淋巴细胞用于细胞融合。在大多数情况下，来自两只小鼠的淋巴结淋巴细胞用于单细胞融合尝试。将卵清蛋白用作抗原，并将融合结果与先前报道的结果（Cell Struct. Funct. 20; 151-156，1995）。使用该方法鉴定了约100个产生目标抗体的阳性威尔斯孔。通过比较，在三次免疫注射后，使用更常规的小鼠脾脏方法鉴定了大约10个阳性威尔斯孔。用注射后14天获得的髂淋巴结淋巴细胞融合后，产生IgM、IgG 1、IgG 2a、IgG 2b和IgG 3的杂交瘤的相对比例分别为14.0%、78.7%、3.2%、3.5%和0.5%。该方法证明了以下优点：（1）单次注射抗原乳剂就足够了，（2）淋巴结在注射后14天即可使用，和（3）获得了高产率的阳性杂交瘤。
A novel method of preparing hybridomas producing mouse monoclonal antibodies was established, called “the mouse iliac lymph node method”. Lymphocytes from enlarged iliac lymph nodes from mice injected intramuscularly at the tail base with an emulsion of antigen and Freund’s adjuvant were used for cell fusion. For the most part, lymph node lymphocytes from two mice were used for a single cell fusion attempt. Ovalbumin was used as the antigen and the results of fusion were compared with the results of a previous report (Cell Struct. Funct. 20; 151–156, 1995). Approximately 100 positive wells producing antibody of interest were identified using this method. By comparison, approximately 10 positive wells were identified using the more conventional mouse spleen method after three immunization injections. The relative proportions of hybridomas producing IgM, IgG1, IgG2a, IgG2b, and IgG3, following fusion using iliac lymph node lymphocytes obtained 14 days after injection were 14.0%, 78.7%, 3.2%, 3.5% and 0.5%, respectively. This method demonstrated the following advantages: (1) a single injection of the antigen emulsion was sufficient, (2) the lymph nodes were ready for use 14 days after injection, and (3) a high yield of positive hybridomas was obtained.
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影响因子：
2.2
作者：
MIRZA, IH;WILKIN, TJ;MOORE, K
通讯作者：
MOORE, K
影响因子：
2.2
作者：
STAHLI, C;STAEHELIN, T;HARING, P
通讯作者：
HARING, P
影响因子：
64.8
作者：
SHULMAN, M;WILDE, CD;KOHLER, G
通讯作者：
KOHLER, G
影响因子：
1.5
作者：
KISHIRO, Y;KAGAWA, M;SADO, Y
通讯作者：
SADO, Y
DOI：
10.1073/pnas.92.20.9348
发表时间：
1995-09-26
影响因子：
11.1
作者：
SPIEKERPOLET, H;SETHUPATHI, P;KNIGHT, KL
通讯作者：
KNIGHT, KL