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[使用不含免疫球蛋白的猪血清和透析组分的杂交瘤培养物]

[Hybridoma cultures using porcine serum not containing immunoglobulin and dialysis fractions].

作者信息

Mantsygin Iu A, Sviatukhina N V, Lisin N M, Morenkov O S

出版信息

Tsitologiia. 1986 Jun;28(6):646-50.

PMID:3529540
Abstract

The whole swine serum was treated with ammonium sulphate to precipitate immunoglobulins. The remained IgG was removed with the use of protein A-sepharose. The hybridoma cells producing monoclonal antibodies to lambda phage (class IgG) were cultured in Dalbecco's modified Eagle medium with addition of a 5% whole swine serum or of a treated unwhole one (final concentration of the protein being 3 mg/ml). Upon these conditions, hybridoma cells had similar growth rate and population density (1-1.3 X 10(6) cells/ml). Maximal antibody concentration was almost similar (80-90 mcg/ml). Purity of a sample of monoclonal antibodies isolated by the method of chromatography with the use of protein A-sepharose from supernatant containing the unwhole serum was no less than 99%, whereas it was considerably lower (12-15%) in the case of the whole serum.

摘要

用硫酸铵处理全猪血清以沉淀免疫球蛋白。使用蛋白A-琼脂糖去除剩余的IgG。产生针对λ噬菌体的单克隆抗体(IgG类)的杂交瘤细胞在添加了5%全猪血清或经处理的非全猪血清(蛋白质终浓度为3mg/ml)的达尔伯克改良伊格尔培养基中培养。在这些条件下,杂交瘤细胞具有相似的生长速率和细胞密度(1-1.3×10⁶个细胞/ml)。最大抗体浓度几乎相似(80-90μg/ml)。通过使用蛋白A-琼脂糖从含有非全猪血清的上清液中进行色谱法分离得到的单克隆抗体样品的纯度不低于99%,而在全猪血清的情况下纯度则低得多(12-15%)。

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