A simple panning method for the selection of cell surface antigen transfectants.
Mouse Ltk- (H-2k) cells that became reactive with anti-H-2d monoclonal antibodies (McAb) after transfection with DNA from H-2d-positive cells were isolated by an indirect panning method. The cells were transfected with DNA from a plasmid containing the selective thymidine kinase (tk) gene in addition to total cell DNA, so that a first selection could be carried out in hypoxanthine/aminopterin/thymidine (HAT) medium. The HAT-selected tk+ transfectants were incubated with anti-H-2d McAb, washed and transferred to dishes coated with purified anti-immunoglobulin (anti-Ig). This 2-step method of panning has the advantage that only the anti-Ig reagent requires purification. Transfected cells clearly reactive with either anti-Ld/Rd or anti-Kd McAb were isolated after only 1 or 2 cycles of panning.
Maryanski
J L
JL
De Plaen
E
E
Van Snick
J
J
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000954:Antigens, Surface / Q000032:analysis*; D000956:Antigens, Viral / Q000032:analysis*; D004279:DNA, Viral / Q000302:isolation & purification; D051379:Mice; D018139:Simplexvirus / Q000235:genetics / Q000276:immunology; D013937:Thymidine Kinase / Q000235:genetics; D014162:Transfection
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D000956:Antigens, Viral; D004279:DNA, Viral; D013937:Thymidine Kinase