Recombinant antibodies to small analytes and prospects for deriving them from synthetic combinatorial libraries
- 1 January 1994
- journal article
- research article
- Published by Taylor & Francis in Food and Agricultural Immunology
- Vol. 6 (3), 277-286
- https://doi.org/10.1080/09540109409354839
Abstract
During the past 4 years, several laboratories have developed new methods of cloning antibody combining site genes from hybridomas or B‐lymphocytes, and functionally expressing them in bacteria, yeast, mammalian cells or plants. At least three research groups have also constructed ‘antibody display’ libraries with vastly diverse sequence permutations of combining site genes in bacteriophage. These semi‐synthetic combinatorial libraries present an antibody repertoire orders of magnitude greater than that accessed by conventional hybridoma technology. They may yield antibodies not obtainable through conventional immunization. We have cloned the immunoglobulin genes from a hybridoma specific for the phenylurea herbicide, diuron, into a phage display vector. The cloned antibody fragments (Fabs) were as sensitive as the parent monoclonal antibody for detecting free diuron in competition enzyme immunoassays. We also derived diuron hapten‐specific clones from synthetic combinatorial phage libraries, but only a few weakly recognized free diuron. The relative merits of deriving Fabs from synthetic phage display libraries, and the steps in engineering new specificities and other properties into recombinant antibodies, are discussed.Keywords
This publication has 39 references indexed in Scilit:
- Direct selection of antibodies that coordinate metals from semisynthetic combinatorial libraries.Proceedings of the National Academy of Sciences, 1993
- Structural repertoire of the human VH segmentsJournal of Molecular Biology, 1992
- Semisynthetic combinatorial antibody libraries: a chemical solution to the diversity problem.Proceedings of the National Academy of Sciences, 1992
- Phage antibodies: will new ‘coliclonal’ antibodies replace monoclonal antibodies?Trends in Biotechnology, 1992
- Assembly of combinatorial antibody libraries on phage surfaces: the gene III site.Proceedings of the National Academy of Sciences, 1991
- Making antibody fragments using phage display librariesNature, 1991
- Combinatorial immunoglobulin libraries on the surface of phage (Phabs): Rapid selection of antigen-specific fabsMethods, 1991
- ‘Sticky feet’-directed mutagenesis and its application to swapping antibody domainsNucleic Acids Research, 1989
- Reshaping the antibody combining site by CDR replacement—tailoring or tinkering to fit?Protein Engineering, Design and Selection, 1988
- Three-Dimensional Structure of AntibodiesAnnual Review of Immunology, 1988