Construction and characterization of a single-chain catalytic antibody.
The antigen-binding (Fab) fragment of the catalytic monoclonal antibody NPN43C9 has recently been cloned by using bacteriophage lambda. By inserting the variable regions of this Fab coding sequence into a (NH2)-VL-linker-VH-(COOH) construct (where VL and VH represent the heavy and light chain variable regions), we have assembled a recombinant gene encoding a catalytic single-chain antigen-binding protein. This protein has been expressed in Escherichia coli and exhibits the same catalytic parameters as the parent monoclonal antibody NPN43C9. Single-chain forms of catalytic antibodies may prove valuable for structural and site-directed mutagenesis studies as well as for large-scale applications of catalytic antibodies.
Gibbs
R A
RA
Department of Chemistry, Pennsylvania State University, University Park 16802.
Posner
B A
BA
Filpula
D R
DR
Dodd
S W
SW
Finkelman
M A
MA
Lee
T K
TK
Wroble
M
M
Whitlow
M
M
Benkovic
S J
SJ
D000595:Amino Acid Sequence; D000906:Antibodies / Q000737:chemistry* / Q000235:genetics; D001483:Base Sequence; D002384:Catalysis; D003001:Cloning, Molecular; D004950:Esterases / Q000235:genetics*; D005822:Genetic Vectors; D007700:Kinetics; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D011994:Recombinant Proteins
D000906:Antibodies; D046911:Macromolecular Substances; D011994:Recombinant Proteins; D004950:Esterases