title: Screening combinatorial antibody libraries for catalytic acyl transfer reactions.
abstract: A bacteriophage lambda vector system for the expression of Fab fragments from the mouse antibody repertoire in Escherichia coli has been described. We have used this system to generate a catalytic antibody from a combinatorial antibody library. Monoclonal antibody 43C9 was raised against a transition state analogue of the hydrolysis of carboxyamide. mRNA from hybridoma cells expressing this antibody was cloned into phage lambda and clones that expressed the mRNA for either the heavy or the light chain of the antibody were isolated. These individual libraries were then crossed to generate a combinatorial library in which clones coexpressed the heavy and light chains. This library was screened for antibodies/Fab fragments that bound to the original antigen with high affinity. DNA sequencing showed that these fragments were the same as those in antibody 43C9. Three different clones were found to catalyse the hydrolysis of carboxyamide. More efficient expression vectors and improved screening techniques should lead to the isolation of many more catalytic antibodies from combinatorial antibody libraries.
authors.affiliation: Department of Molecular Biology & Chemistry, Research Institute of Scripps Clinic, La Jolla, CA 92037.
authors.forename: L
authors.initials: L
authors.lastname: Sastry
authors.forename: M
authors.initials: M
authors.lastname: Mubaraki
authors.forename: K D
authors.initials: KD
authors.lastname: Janda
authors.forename: S J
authors.initials: SJ
authors.lastname: Benkovic
authors.forename: R A
authors.initials: RA
authors.lastname: Lerner
mesh_terms: D000215:Acylation; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000378:metabolism*; D010582:Bacteriophage lambda; D002384:Catalysis; D005822:Genetic Vectors; D015698:Genomic Library; D007140:Immunoglobulin Fab Fragments / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D011994:Recombinant Proteins