High-performance affinity isolation of lymphocyte membrane receptors on biotinylated antigen and avidin-coated beads.
Isolation of lymphocyte membrane receptors can be achieved by high-performance liquid chromatography using immobilized streptavidin as the ligand and biotinylated antigen. Activated lymphocytes were allowed to react with biotin-labelled antigen prior to harvesting. The cells were disrupted and their membranes solubilized before passing the suspension through the avidin affinity column. The biotinylated antigen acted as an efficient receptor probe, which helped to maintain the integrity of the receptor during the isolation procedure. The biotin also acted as the substrate that attaches to the immobilized avidin. Recovery of the bound receptor was achieved by dissociation of the receptor from the antigen and recovery of the receptor in the effluent during the elution phase of the separation.
Phillips
T M
TM
George Washington University Medical Center, Washington, DC 20037.
Frantz
S C
SC
Babashak
J V
JV
Chmielinska
J J
JJ
D001360:Avidin; D001710:Biotin; D002846:Chromatography, Affinity; D008214:Lymphocytes / Q000276:immunology*; D011971:Receptors, Immunologic / Q000302:isolation & purification*
D011971:Receptors, Immunologic; D001360:Avidin; D001710:Biotin