Affinity purification of latex antigens.
Latex extracts are complex mixtures of antigenic peptides. We attempted to raise monoclonal antibodies to latex and to use these antibodies to purify latex antigens. A monoclonal antibody, CRI-C, was raised by standard techniques. Peptides of nonammoniated latex (NAL) and ammoniated latex were electrophoretically separated and transferred for immunoblots. CRI-C was covalently attached to an agarose column. NAL was passed over the column, and purified antigen was then eluted. The eluate was analyzed by sodium dodecylsulfate-polyacrylamide gel electrophoresis and RAST inhibition with sera from health care workers and children with spina bifida. CRI-C recognized a single band in ammoniated latex immunoblots and several distinct bands in NAL. The affinity-purified antigen of CRI-C (C-Ag) had multiple bands of less than 20 kd and was 3.9 times more potent in RAST inhibition than NAL when sera from patients with spina bifida were used. However, when health care workers' sera were used, there was no significant difference in the inhibitory potency of NAL and C-Ag. CRI-C appears to recognize a distinct and important epitope in the IgE immune response to latex of patients with spina bifida.
Slater
J E
JE
Center for Cancer and Transplantation Biology, Children's Research Institute, Washington, D.C.
Trybul
D E
DE
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000941:Antigens / Q000276:immunology* / Q000302:isolation & purification*; D002675:Child, Preschool; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D006967:Hypersensitivity / Q000276:immunology; D015151:Immunoblotting; D007073:Immunoglobulin E / Q000276:immunology; D007840:Latex; D051379:Mice; D008807:Mice, Inbred BALB C; D011852:Radioallergosorbent Test
D000911:Antibodies, Monoclonal; D000941:Antigens; D007840:Latex; D007073:Immunoglobulin E