ZZ polyester beads: an efficient and simple method for purifying IgG from mouse hybridoma supernatants.
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.
Lewis
John G
JG
Steroid and Immunobiochemistry Laboratory, Canterbury Health Laboratories, Christchurch, New Zealand. john.lewis@cdhb.govt.nz
Rehm
Bernd H A
BH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000302:isolation & purification*; D001665:Binding Sites; D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006825:Hybridomas / Q000378:metabolism*; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000302:isolation & purification*; D051379:Mice; D054813:Polyhydroxyalkanoates / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013205:Staphylococcal Protein A / Q000235:genetics / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D054813:Polyhydroxyalkanoates; D011994:Recombinant Proteins; D013205:Staphylococcal Protein A