Jones S L, Cox J C, Pearson J E
Commonwealth Serum Laboratories, Parkville, Australia.
J Immunol Methods. 1990 May 25;129(2):227-31. doi: 10.1016/0022-1759(90)90443-y.
The production of monoclonal antibodies is conveniently achieved by propagation of antibody-secreting hybridoma cells as ascites tumours in mice. With a view to improve production and minimise animal usage, the choice of primer (pristane or Freund's incomplete adjuvant (FIA] and the effect of sex of mouse (female vs. uncastrated male) upon antibody yields were compared. The volume of ascitic fluid harvested from 134 FIA-primed mice was 697 ml, which was significantly (P less than 0.02) greater than the 343 ml collected from 116 pristane-primed mice. Moreover, FIA-primed mice required tapping for only 11 days (90% in 7 days) compared to the 17 days needed for pristane-primed mice. While there was little difference between the mean IgG1 concentration (as determined by enzyme immunoassay) of both ascites pools (4.9 mg/ml compared to 4.6 mg/ml), the volume differences equated to yields of 25.5 mg/mouse and 13.5 mg/mouse respectively. The FIA-primed mice were also shown to produce on average more (P less than 0.02) ascites per day. There was no significant difference found between the yield of monoclonal antibodies obtained from either uncastrated males (22.1 mg/mouse) or females (23.1 mg/mouse). The use of FIA to prime mice for the in vivo production of monoclonal antibodies should improve the yield to over 20 mg/mouse, thereby reducing the number of mice required.
通过在小鼠体内将分泌抗体的杂交瘤细胞作为腹水肿瘤进行繁殖,可方便地生产单克隆抗体。为了提高产量并尽量减少动物使用量,比较了引物( pristane或弗氏不完全佐剂(FIA))的选择以及小鼠性别(雌性与未阉割雄性)对抗体产量的影响。从134只经FIA预处理的小鼠中收集的腹水量为697毫升,这显著(P小于0.02)大于从116只经pristane预处理的小鼠中收集的343毫升。此外,经FIA预处理的小鼠只需抽取11天(7天内90%),而经pristane预处理的小鼠需要17天。虽然两个腹水样本的平均IgG1浓度(通过酶免疫测定法测定)之间差异不大(分别为4.9毫克/毫升和4.6毫克/毫升),但体积差异分别相当于每只小鼠产生25.5毫克和13.5毫克。经FIA预处理的小鼠平均每天产生的腹水也更多(P小于0.02)。从未阉割雄性(22.1毫克/小鼠)或雌性(23.1毫克/小鼠)获得的单克隆抗体产量之间没有显著差异。使用FIA预处理小鼠用于体内生产单克隆抗体应能将产量提高到每只小鼠超过20毫克,从而减少所需小鼠的数量。