Harnessing malE for the study of antigen/antibody recognitions.
The construction of hybrids between the variable fragment (Fv) of antibodies and protein MalE of Escherichia coli at the genetic level makes possible their preparation in a functional state, independently of any interaction with the antigen. We used such hybrids and a mutagenesis approach to study the recognition between antibody D1.3 and its antigen lysozyme, and its maturation. We subsequently transformed D1.3 into a reagentless fluorescent biosensor by knowledge-based design.
Bedouelle
Hugues
H
Department of Structural Biology and Chemistry, CNRS URA 2185, Institut Pasteur, Paris, France. hbedouel@pasteur.fr
Renard
Martial
M
Belkadi
Laurent
L
England
Patrick
P
D000818:Animals; D000937:Antigen-Antibody Reactions / Q000276:immunology*; D015374:Biosensing Techniques; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D007128:Immunoglobulin Fragments / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D033902:Periplasmic Binding Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000276:immunology
D029968:Escherichia coli Proteins; D007128:Immunoglobulin Fragments; C461619:MalE protein, E coli; D033902:Periplasmic Binding Proteins; D011993:Recombinant Fusion Proteins; C055966:immunoglobulin Fv