title: Fully in vitro selection of recombinant antibodies.
abstract: Antibodies are essential for the identification and characterization of proteins. In the current postgenomic era the need for highly specific antibodies has further increased not only for research applications but also because they represent one of the most promising therapeutic options, especially in the field of cancer treatment. One appealing approach for rapid and inexpensive antibody generation is the use of phage display. This technique allows for a fast and animal-free selection of highly functional alternatives to classical antibodies. However, one strong limitation of this recombinant approach has been the difficulty in producing and purifying antigens. These steps have to be adjusted for each new target, are time consuming and sometimes present an insurmountable obstacle. Here we report the development of new antibody selection approach where antigens are produced through in vitro translation and are used directly and without the need for purification. With this approach we were able to rapidly select recombinant antibodies directed against GFP and the mammalian protein tsg101, respectively. We believe that our method greatly facilitates antigen preparation and thus may broaden the use of the recombinant approach for antibody generation, especially since the technique could in the future be adapted to a high-throughput technology, thus further accelerating antibody selection.
authors.affiliation: Transfer Department, Institut Curie, Paris, France.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Moutel
authors.forename: Ole
authors.initials: O
authors.lastname: Vielemeyer
authors.forename: Hulin
authors.initials: H
authors.lastname: Jin
authors.forename: Séverine
authors.initials: S
authors.lastname: Divoux
authors.forename: Philippe
authors.initials: P
authors.lastname: Benaroch
authors.forename: Franck
authors.initials: F
authors.lastname: Perez
mesh_terms: D000906:Antibodies / Q000235:genetics* / Q000378:metabolism; D000941:Antigens / Q000378:metabolism; D001710:Biotin / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D056827:Endosomal Sorting Complexes Required for Transport; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000235:genetics* / Q000378:metabolism; D008856:Microscopy, Fluorescence; D019151:Peptide Library; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D000099177:Tsg101 Protein
chemical_list: D000906:Antibodies; D000941:Antigens; D001710:Biotin; D004268:DNA-Binding Proteins; D056827:Endosomal Sorting Complexes Required for Transport; D049452:Green Fluorescent Proteins; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D011994:Recombinant Proteins; D014157:Transcription Factors; D000099177:Tsg101 Protein