Isolation of monoclonal antibodies recognizing rare and dominant epitopes in plant vascular cell walls by phage display subtraction.
A combination of phage display antibody technology and a subtraction method provides a powerful tool for the isolation of novel biomarkers. However, the dilemma that high stringency screening often reduces the diversity in the subtracted phage antibody repertoires and that it is difficult to isolate phage antibody against rare epitopes remain. Therefore, we carefully monitored the occupancy of differentiation-specific clones in a phage antibody library through an enrichment process, and succeeded in isolating monoclonal antibodies against rare and dominant epitopes in plant vascular cell walls. We also report that clones with stop and frameshift mutations significantly survived the enrichment process, owing to noncanonical translation mechanisms.
Department of Biological Sciences, Graduate School of Science, University of Tokyo, Hongo, Tokyo, Japan. shino@biol.s.u-tokyo.ac.jp
Naoki
N
Shinohara
Hiroo
H
Fukuda
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D019659:Asteraceae; D017104:Bacteriophage M13 / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D002454:Cell Differentiation / Q000235:genetics / Q000276:immunology; D002473:Cell Wall / Q000235:genetics / Q000378:metabolism*; D002999:Clone Cells; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D016056:Immunodominant Epitopes / Q000235:genetics / Q000378:metabolism; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D019151:Peptide Library; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
D000911:Antibodies, Monoclonal; D016056:Immunodominant Epitopes; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D010940:Plant Proteins