Lewis John G, Rehm Bernd H A
Steroid and Immunobiochemistry Laboratory, Canterbury Health Laboratories, Christchurch, New Zealand.
J Immunol Methods. 2009 Jul 31;346(1-2):71-4. doi: 10.1016/j.jim.2009.04.011. Epub 2009 Apr 24.
ZZ polyester beads are polyhydroxyalkanoate granules which display, at high density, the ZZ domain of protein A as a fusion protein covalently linked to the polyester core. These granules are produced directly, in one step, by recombinant E. coli. We have used these granules produced in a one step process to purify mouse IgG1, IgG2a and IgG2b from mouse hybridoma supernatants. Purified immunoglobulins were analysed by enzyme-linked immonosorbent assay, agarose gel electrophoresis and SDS-PAGE. The results showed that the recovery of IgG is 70% or greater with a significant degree of purity. ZZ polyester beads hence offer a rapid and novel method to purify IgG from mouse hybridoma culture supernatants. As a negative control we used culture supernatant from an IgM secreting hybridoma and showed that it did not bind to ZZ polyester beads.
ZZ聚酯珠是聚羟基链烷酸酯颗粒，其以高密度展示作为与聚酯核心共价连接的融合蛋白的蛋白A的ZZ结构域。这些颗粒由重组大肠杆菌一步直接产生。我们已经使用通过一步法产生的这些颗粒从小鼠杂交瘤上清液中纯化小鼠IgG1、IgG2a和IgG2b。通过酶联免疫吸附测定、琼脂糖凝胶电泳和SDS-PAGE对纯化的免疫球蛋白进行分析。结果表明，IgG的回收率为70%或更高，且纯度相当高。因此，ZZ聚酯珠提供了一种从小鼠杂交瘤培养上清液中纯化IgG的快速且新颖的方法。作为阴性对照，我们使用了分泌IgM的杂交瘤的培养上清液，并表明它不与ZZ聚酯珠结合。
