Perfusion immunoaffinity chromatography and its application in analysis and purification of biomolecules.
Immunoaffinity cartridges have been prepared by immobilizing the monoclonal antibody to bovine growth hormone (bGH) and bovine growth hormone releasing factor (bGRF) on the streptavidin-coated perfusion media. The cartridge-immobilized antibody to bGH has been used for the analysis of standard sample of recombinant bGH (r-bGH). The cartridge immobilized bGRF has been applied for purification of the C-and N-terminal antibodies and their conjugates with horseradish peroxidase. The purity of the antibody fractions purified by bGRF cartridges has been tested by capillary zone electrophoresis. It has been shown that the immunoaffinity cartridges prepared by immobilizing the antigen offer more biological specificity for the purification of antibody than the Protein G cartridges. The stability and life time of the immunoaffinity cartridges are mainly dependent on the stability of the biomolecules immobilized on them. Immunoaffinity cartridges based on the perfusion media offer us the tools for rapid analysis and purification of antigens or antibodies as well as for determination of the biological activity between them at a very low back pressure in the columns.
Zou
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H
National Chromatographic R. and A. Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, China.
Zhang
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Y
Lu
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P
Krull
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IS
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000302:isolation & purification*; D000941:Antigens / Q000737:chemistry / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000737:chemistry; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods*; D004355:Drug Stability; D019075:Electrophoresis, Capillary / Q000379:methods; D013006:Growth Hormone / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D013007:Growth Hormone-Releasing Hormone / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D006735:Horseradish Peroxidase / Q000032:analysis / Q000737:chemistry; D007124:Immunoenzyme Techniques; D051379:Mice; D009419:Nerve Tissue Proteins / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D013056:Spectrophotometry, Ultraviolet; D019809:Streptavidin
D000911:Antibodies, Monoclonal; D000941:Antigens; D001426:Bacterial Proteins; C029152:G-substrate; D009419:Nerve Tissue Proteins; D011994:Recombinant Proteins; D013006:Growth Hormone; D019809:Streptavidin; D013007:Growth Hormone-Releasing Hormone; D006735:Horseradish Peroxidase