ZZ polyester beads: An efficient and simple method for purifying IgG from mouse hybridoma supernatants

Abstract

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.