Federspiel G, McCullough K C, Kihm U
Institut für Viruskrankheiten und Immunprophylaxe (IVI), Basel, Switzerland.
J Immunol Methods. 1991 Dec 15;145(1-2):213-21. doi: 10.1016/0022-1759(91)90329-e.
Six hybridoma clones (three producing anti-bovine viral diarrhoea antibodies and three producing anti-hog cholera virus antibodies) were studied with respect to their growth in type II serum-free medium. This was found to be successful in all cases when the serum-free medium was supplemented with 1% v/v Nutridoma, but only after adaptation over a small number of subcultures. Adapted hybridoma cultures could be grown in 250 ml spinner flasks, and the antibody-containing supernatants concentrated by tangential flow filtration. These were compared both with conventional cultures of the clones grown in serum-containing media and with ascites fluids generated using the latter type of culture. Adaptation to growth in the Nutridoma-based serum-free medium did not impair the capacity of the cells to produce and secrete antibodies, although their growth rate was slightly reduced when 250 ml spinner cultures were employed. The titre obtained from the ascites fluids were, as expected, higher than those from the tissue cultures, but contamination by other proteins, a significant problem in ascites fluids, did not present itself with the serum-free cultures. Consequently, 'serum-free' hybridoma antibodies could easily be concentrated in a relatively pure form, with respect to immunoglobulins present, having no need for affinity purification procedures. In addition, the acute in vivo problems of solid tumours and haemorrhage production could be avoided through the use of in vitro methods.
研究了六个杂交瘤克隆（三个产生抗牛病毒性腹泻抗体，三个产生抗猪霍乱病毒抗体）在II型无血清培养基中的生长情况。当无血清培养基中添加1%（v/v）的纽蛋白时，发现在所有情况下这都是成功的，但这仅在经过少量传代适应后才实现。适应后的杂交瘤培养物可以在250毫升转瓶中生长，并且含有抗体的上清液通过切向流过滤进行浓缩。将这些与在含血清培养基中生长的克隆的传统培养物以及使用后一种培养物产生的腹水进行了比较。适应在基于纽蛋白的无血清培养基中生长并没有损害细胞产生和分泌抗体的能力，尽管当采用250毫升转瓶培养时其生长速率略有降低。如预期的那样，从腹水中获得的滴度高于从组织培养物中获得的滴度，但是腹水的一个重大问题——其他蛋白质的污染，在无血清培养物中并未出现。因此，就存在的免疫球蛋白而言，“无血清”杂交瘤抗体可以很容易地以相对纯的形式浓缩，无需亲和纯化程序。此外，通过使用体外方法可以避免实体瘤和出血产生的急性体内问题。
