ZZ polyester beads: An efficient and simple method for purifying IgG from mouse hybridoma supernatants
ZZ polyester beads: An efficient and simple method for purifying IgG from mouse hybridoma supernatants
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DOI：
10.1016/j.jim.2009.04.011
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发表时间：
2009-07-31
影响因子：
2.2
通讯作者：
Rehm, Bernd H. A.
中科院分区：
文献类型：
--
作者：
Lewis, John G.;Rehm, Bernd H. A.
ZZ聚酯微珠是聚羟基烷酸颗粒，它以高密度显示蛋白A的ZZ结构域，作为共价连接到聚酯核心的融合蛋白。这些颗粒是由重组大肠杆菌一步直接生产的。我们用一步法生产的这些颗粒从小鼠杂交瘤上清液中纯化了小鼠IgG1、IgG2a和IgG2b。用酶联免疫吸附试验、琼脂糖凝胶电泳法和SDS-PAGE对纯化的免疫球蛋白进行分析。结果表明，免疫球蛋白的回收率在70%以上，纯度较高。因此，ZZ聚酯小球为从小鼠杂交瘤培养上清液中纯化免疫球蛋白提供了一种快速、新颖的方法。作为阴性对照，我们使用了一株分泌IgM的杂交瘤的培养上清，结果表明它不与ZZ聚酯小球结合。(C)2009爱思唯尔B.V.保留所有权利。
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. (c) 2009 Elsevier B.V. All rights reserved.