Affinity selection of cDNA libraries by lambda phage surface display.
Bacteriophage lambda surface display was used to isolate cDNA clones encoding autoantigens recognized by sera from patients with Sjögren's syndrome (SS). We made cDNA libraries from human HeLa and HepG2 cells, using the expression vector lambdafoo. By repeating affinity selection of the libraries with the sera immobilized in microtiter wells, we isolated three clones that encode previously unknown antigens as well as four clones previously known as SS autoantigens. The newly identified autoantigens include TRK-fused gene product (TFG), survival motor neuron gene product (SMN) and pM5, which has a similarity to the metal-binding domain of human fibroblast collagenase. Thus, the bacteriophage lambda surface display is powerful for isolating cDNA clones by affinity screening.
Niwa
M
M
Department of Cell Biology, The Scripps Research Institute, 10550 N. Torrey Pines Rd., La Jolla, CA 92037, USA.
Maruyama
H
H
Fujimoto
T
T
Dohi
K
K
Maruyama
I N
IN
D000915:Antibody Affinity; D001324:Autoantigens / Q000235:genetics* / Q000276:immunology; D010582:Bacteriophage lambda / Q000235:genetics; D001667:Binding, Competitive; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics*; D015723:Gene Library; D006367:HeLa Cells; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D017422:Sequence Analysis, DNA; D014407:Tumor Cells, Cultured
D001324:Autoantigens; D018076:DNA, Complementary; D007106:Immune Sera