Pakkanen R, Kanttinen A, Satama L, Aalto J
Valio Bioproducts Ltd., Turku Technology Centre, Finland.
Appl Microbiol Biotechnol. 1992 Jul;37(4):451-6. doi: 10.1007/BF00180967.
Fractions of bovine colostrum were prepared and their ability to support the growth of mouse-mouse hybridomas in culture was tested. Whey was prepared from defatted colostrum by removal of casein using acid precipitation. An ultrafiltrate was obtained from cleared whey by filtration through membranes with a nominal molecular mass cut-off of 100,000 Da. Colostrum ultrafiltrate contained 1.16 milligrams protein, 0.24 milligrams immunoglobulin G (IgG) and less than 0.24 EU (endotoxin unit)/ml endotoxins. The effect of defatted colostrum, whey and ultrafiltrate as serum substitutes was examined by cultivation of hybridoma cells in minimal essential medium containing different concentrations of the supplements. Under optimal conditions in ultrafiltrate-supplemented medium, the maximal cell concentration was 35-40% of that obtained using 10% foetal bovine serum, and IgG production per cell was equal to that achieved using serum. In 1% defatted colostrum the maximum hybridoma concentration was about 30% of that in 10% serum, but at higher concentrations hybridoma growth was significantly reduced. The growth-promoting activity of whey was low. The results show that bovine colostrum ultrafiltrate provides a very attractive alternative to serum for production of monoclonal antibodies.
制备了牛初乳的各个组分,并测试了它们在培养中支持小鼠 - 小鼠杂交瘤生长的能力。通过酸沉淀去除酪蛋白,从脱脂初乳中制备乳清。通过用标称分子量截留值为100,000 Da的膜过滤,从澄清的乳清中获得超滤物。初乳超滤物含有1.16毫克蛋白质、0.24毫克免疫球蛋白G(IgG)和每毫升少于0.24 EU(内毒素单位)的内毒素。通过在含有不同浓度补充剂的最低必需培养基中培养杂交瘤细胞,研究了脱脂初乳、乳清和超滤物作为血清替代品的效果。在补充超滤物的培养基的最佳条件下,最大细胞浓度为使用10%胎牛血清时获得的最大细胞浓度的35 - 40%,并且每个细胞的IgG产量与使用血清时相当。在1%脱脂初乳中,杂交瘤的最大浓度约为10%血清中的30%,但在更高浓度下杂交瘤生长显著降低。乳清的促生长活性较低。结果表明,牛初乳超滤物为单克隆抗体的生产提供了一种非常有吸引力的血清替代品。