Molecules with antibody combining sites that catalyze hydrolysis reactions through use of a charged hapten

Monoclonal antibody molecules or molecules containing antibody combining site portions that catalytically hydrolyze a preselected carboxylic acid ester bond of a reactant ligand, the antibody combining site of said molecules binding to:

• (a) a reactant ligand containing the preselected carboxylic acid ester bond that is hydrolyzed, said reactant ligand being represented by the structure ##STR19## wherein R.sup.1 and R.sup.2 represent carbon atom-containing chemical residues of the reactant, and
• --X--is --O--;
• and
• (b) a haptenic ligand that is structurally analogous to said reactant ligand, said haptenic ligand being represented by the structure ##STR20## where R.sup.1' and R.sup.2' represent carbon atom-containing residues that are structurally analogous to R.sup.1 and R.sup.2, respectively, and at least one of R.sup.1' and R.sup.2' containing a group bearing an ionic charge in aqueous solution at physiological pH values, at least one said group providing ionic charge at physiological pH values being located within a spherical volume defined by a radius of about 7 .ANG.ngstroms from said ##STR21## group of said structure, and R.sup.3' is H.

A method of preparing cells that when cultured in a medium produce antibody molecules or molecules containing antibody combining site portions that catalytically hydrolyze a preselected carboxylic acid ester bond of a reactant ligand comprising the steps of:

• (a) immunizing an animal with an immunogen that includes a haptenic ligand that contains a tetrahedral carbon atom bonded to a hydroxyl group as well as to a saturated carbon atom at a position in the haptenic ligand corresponding to the position of the carbonyl group as well as to the carbonyl-bonded heteroatom, respectively, of the preselected carboxylic acid ester bond to be hydrolyzed, said haptenic ligand further including a group bearing an ionic charge in aqueous solution at physiological pH values, the ionic charge-bearing group being absent from a corresponding position of said reactant ligand and located within a spherical volume defined by a radius of about 7 .ANG.ngstroms from said tetrahedral carbon atom;
• (b) maintaining said animal for a time period sufficient for said animal to secrete antibodies that immunoreact with said haptenic ligand;
• (c) transferring genes that encode antibody molecules or molecules containing antibody combining site portions from antibody-producing cells of said maintained, immunized animal of step (b) into host cells to form hybrid cells that contain genes from at least two sources, and which formed hybrid cells (i) produce antibody molecules or molecules containing antibody combining site portions from said transferred genes when cultured and (ii) can be cultured substantially indefinitely;
• (d) culturing the hybrid cells in an appropriate culture medium for a time period sufficient for those hybrid cells to produce antibody molecules or molecules containing antibody combining site portions;
• (e) recovering antibody molecules or molecules containing antibody combining site portions from the cultured hybrid cells;
• (f) screening the obtained antibody molecules or molecules containing antibody combining site portions to identify a hybrid cell that produces antibody molecules or molecules containing antibody combining site portions that catalytically hydrolyze said predetermined carboxylic acid ester bond;
• and
• (g) growing clones of said identified hybrid cell that produces antibody molecules or molecules containing antibody combining site portions that catalytically hydrolyze said predetermined carboxylic acid ester bond.