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首页> 外文期刊>The Kasetsart Journal >Production of Fish Serum Products as Substitute for Fetal Bovine Serum in Hybridoma Cell Cultures from Surimi Industrial Waste
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Production of Fish Serum Products as Substitute for Fetal Bovine Serum in Hybridoma Cell Cultures from Surimi Industrial Waste

机译：鱼糜产品替代鱼糜工业废料在杂交瘤细胞培养中替代胎牛血清

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摘要

The hybridoma cell culture procedures mostly utilize fetal bovine serum (FBS) to supplement the culture media, which contributes to the high cost of production of monoclonal antibody. Previous research has shown interesting results on the utilization of fish serum (FS) prepared from wash water of surimi processing line. In this study, FS was used as a substitute for FBS in two types of hybridoma cell cultures. Cell viability was measured using MTT colorant while IgG monoclonal antibody secretion was measured using ELISA method. SDS electrophoresis was run to observe FS protein profiles. Nitrogenous compounds of FS were also determined. Based on the results, it was found that FS prepared from surimi factory wash water contained proteins with molecular weights ranging from 66 to 12 kD. The serum contained TVN and TMA, signalling the need to improve the washing process in the factory, particulary the waste water treatment in formulating the fish serum. The two hybridomas secreting IgG anti-chikungunya (105) and anti-dengue virus (H2) grew differently in the FS-substituted medium, where the 105 hybridoma showed less cell growth. The cells in all substituted cultures produced antibody until the third passage; however, substitution of 2% FBS with FS resulted in the higher antibody secretion, especially in cultures maintained for 96 hours after the third passage.
机译：杂交瘤细胞培养程序主要利用胎牛血清（FBS）来补充培养基，这导致生产单克隆抗体的成本较高。先前的研究显示了利用鱼糜加工线洗涤水制备的鱼浆（FS）的有趣结果。在这项研究中，FS在两种类型的杂交瘤细胞培养物中用作FBS的替代品。使用MTT着色剂测量细胞活力，同时使用ELISA方法测量IgG单克隆抗体的分泌。进行SDS电泳以观察FS蛋白谱。还确定了FS的含氮化合物。根据该结果，发现由鱼糜工厂洗涤水制备的FS包含分子量为66至12kD的蛋白质。血清中含有TVN和TMA，表明需要改进工厂的洗涤过程，尤其是配制鱼血清时的废水处理。分泌IgG抗基孔肯雅热（105）和抗登革热病毒（H2）的两种杂交瘤在FS替代培养基中的生长不同，其中105杂交瘤的细胞生长较少。所有替代培养物中的细胞都产生抗体，直到第三次传代为止。但是，用FS替代2％FBS会导致更高的抗体分泌，特别是在第三次传代后维持96小时的培养物中。

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