Isolation of a specific membrane protein by immunoaffinity chromatography with biotinylated antibodies immobilized on avidin-coated glass beads.
Avidin-coated, solid glass beads have been used as an immobilization support for attaching biotinylated antibodies. These beads have been packed into analytical, semi-preparative and preparative columns and used to isolate the B27 histocompatibility anigen (HLA) from human lymphocytes. The beads provided a suitable column material for all three chromatographic procedures and, depending on the size of the immunoaffinity column, B27 antigen could be isolated in nanogram to microgram quantities. Polyacrylamide gel electrophoresis demonstrated the presence of only a single band in the immunoaffinity peaks isolated by all three procedures. Enzyme-linked immunosorbent assay analysis of these immunoaffinity-isolated materials revealed that they were biologically active and could be used to determine the levels of anti-B27 antibodies in clinical studies.
Babashak
J V
JV
Kontes Scientific Glassware, Vineland, NJ.
Phillips
T M
TM
D000911:Antibodies, Monoclonal; D001360:Avidin; D001710:Biotin; D002846:Chromatography, Affinity / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005898:Glass; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism; D008565:Membrane Proteins / Q000276:immunology / Q000302:isolation & purification*; D013056:Spectrophotometry, Ultraviolet
D000911:Antibodies, Monoclonal; D008565:Membrane Proteins; D001360:Avidin; D001710:Biotin