Production and characterization of a monoclonal antibody against Schistosoma japonicum glutathione S-transferase.
Schistosoma japonicum glutathione S-transferase (GST), expressed from a pGEX plasmid, was isolated from Escherichia coli cells and used to immunize mice in order to generate specific anti-GST monoclonal antibodies. Using a modified immunization and fusion procedure, one stable hybridoma clone secreting an anti-GST antibody (alpha GST-1) was obtained. Milligram quantities of this antibody were produced in vitro in a miniPERM bioreactor and subsequently purified by protein G affinity chromatography. The characteristics of this antibody were investigated by enzyme-linked immunosorbent assays and immunoblotting experiments. The alpha GST-1 antibody was found to react specifically with GST and GST fusion proteins and demonstrated no reactivity with normal E. coli proteins. This monoclonal antibody should be a valuable reagent for tracing the production of GST fusion proteins and possibly for affinity purification of GST fusion proteins.
Campbell
M J
MJ
Department of Surgery, Stanford University School of Medicine, CA 94305, USA.
McFall
P
P
Niederhuber
J E
JE
D000818:Animals; D000909:Antibodies, Helminth / Q000096:biosynthesis* / Q000737:chemistry; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000737:chemistry; D002846:Chromatography, Affinity; D005982:Glutathione Transferase / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry; D012549:Schistosoma japonicum / Q000201:enzymology* / Q000276:immunology*
D000909:Antibodies, Helminth; D000911:Antibodies, Monoclonal; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase