Department of Biochemical Science and Technology, Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima, 890-0065, Japan,
Cytotechnology. 2000 Jul;33(1-3):21-6. doi: 10.1023/A:1008163312841.
We compared serum and protein-free cultures ofa ras-amplified recombinant BHK-21 cell line(ras-rBHK-IgG), which hyperproduces a lungcancer specific recombinant human monoclonal antibody. Ras-rBHK-IgG cells were shown to grow well, evenin protein-free medium and to be morphologicallysimilar to cells cultured in serum containing medium. However, the growth rate of ras-rBHK-IgG cellswas considerably slower in protein-free medium, whichresults in a longer maintenance period compared with cells cultured in serum containing medium. In addition, it was found that antibody production in protein-free culture had a ten times higher maximum than cells cultured in serum containing medium. On theother hand, in high density culture, using the hollowfiber bioreactor system, ras-rBHK-IgG cellscould be maintained for a month in protein-freeculture in contrast with serum culture, which onlylasted for half a month. However, the markedincrease of antibody production was not observed. A total amount of about 15 mg of the recombinantantibody, obtained in protein-free culture, was abouttwo times of that obtained in serum culture, and wasshown to be reactive to lung cancer cells in tissue. From these properties in protein-free medium, it isconcluded that protein-free culture of ras-rBHK-IgG cells is suitable for middle scaleproduction of recombinant human monoclonal antibody.
我们比较了一株 ras 扩增的重组 BHK-21 细胞系（ras-rBHK-IgG）的血清和无蛋白培养物，该细胞系高表达一种肺癌特异性的重组人源单克隆抗体。ras-rBHK-IgG 细胞在无蛋白培养基中生长良好，形态与含血清培养基中培养的细胞相似。然而，ras-rBHK-IgG 细胞在无蛋白培养基中的生长速度要慢得多，与含血清培养基中培养的细胞相比，维持时间更长。此外，发现在无蛋白培养物中的抗体产生比在含血清培养基中培养的细胞高 10 倍。另一方面，在高密度培养中，使用中空纤维生物反应器系统，ras-rBHK-IgG 细胞在无蛋白培养中可以维持一个月，而在血清培养中只能维持半个月。然而，并没有观察到抗体产生的显著增加。从无蛋白培养物中获得的约 15 毫克重组抗体，是从血清培养物中获得的两倍左右，并且在组织中显示出对肺癌细胞的反应性。从无蛋白培养基中的这些特性来看，结论是 ras-rBHK-IgG 细胞的无蛋白培养适合于重组人源单克隆抗体的中规模生产。
