Preparation of sheep and cattle immunoglobulins with antibody activity by high-performance liquid chromatography.
High-performance anion-exchange chromatography (HPIEC) and high-performance size-exclusion chromatography (HPSEC) were used to purify serum IgM and IgG from sheep and cattle. Pooled serum from normal cattle and sheep and serum from sheep, infected with two different viruses, were prepared for HPIEC by chromatography on CM-Affi-Gel Blue. After HPIEC, fractions containing IgG and IgM were pooled and concentrated and further purified by HPSEC. The purity of fractions from HPIEC and HPSEC were evaluated by immunoelectrophoresis, protein-A-Sepharose affinity chromatography, and sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Monoclonal antibody specific for bovine IgG2 was used to assay for IgG2 and IgG2 contamination of other fractions. The antibody activity to ovine adenovirus 5 and to ovine progressive pneumonia virus was assayed by neutralization and immunodiffusion. Antibody activity was retained against both viruses in the fractions containing IgG1 and IgM. This high-performance liquid chromatography procedure was a rapid preparative method to produce specific immunoglobulins and could be used to evaluate the purity of immuno-reagents.
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification*; D000914:Antibodies, Viral / Q000302:isolation & purification; D002417:Cattle; D002846:Chromatography, Affinity; D002848:Chromatography, DEAE-Cellulose; D002851:Chromatography, High Pressure Liquid; D007122:Immunoelectrophoresis; D007074:Immunoglobulin G / Q000302:isolation & purification; D007075:Immunoglobulin M / Q000302:isolation & purification; D007136:Immunoglobulins / Q000302:isolation & purification*; D011863:Radioimmunoassay; D012756:Sheep; D012967:Sodium Dodecyl Sulfate; D014780:Viruses / Q000276:immunology
D000906:Antibodies; D000914:Antibodies, Viral; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D007136:Immunoglobulins; D012967:Sodium Dodecyl Sulfate