Eupergit C as a carrier for high-performance liquid chromatographic-based immunopurification of antigens and antibodies.
An immunoaffinity purification system using C30N and C1Z Eupergit C beads was developed and optimized. Poly- and monoclonal antibodies were purified using immobilized antigens and antigens were purified using immobilized antibodies. Antigens were used that possess enzymic activities and the efficiency of antigen binding was determined from the enzymic activity of the matrix-bound immunocomplexes. High-performance immunoaffinity purification using Eupergit C beads proved to be highly specific, reproducible, free from protein leakage and possessed a low degree of non-specific adsorption of tissue proteins. These characteristics of the system were illustrated by the isolation of immunoglobulin G from serum and of human decidua proteins from the decidua tissue and from seminal plasma. These proteins were obtained at high purity in a single purification step, as shown by sodium dodecyl sulphate-polyacrylamide gel electrophoresis.
Department of Biotechnology, George Wise Faculty of Life Sciences, Tel-Aviv University, Israel.
G
G
Fleminger
T
T
Wolf
E
E
Hadas
B
B
Solomon
D000906:Antibodies / Q000302:isolation & purification*; D000941:Antigens / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid; D003656:Decidua / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification; D008024:Ligands; D008297:Male; D011108:Polymers; D011506:Proteins / Q000032:analysis; D012661:Semen / Q000032:analysis; D012967:Sodium Dodecyl Sulfate
D000906:Antibodies; D000941:Antigens; D007074:Immunoglobulin G; D008024:Ligands; D011108:Polymers; D011506:Proteins; D012967:Sodium Dodecyl Sulfate; C065843:Eupergit C