Patent ID: 8445642

Claim:
A method for preparing monoclonal antibodies that are capable of discriminating between at least two stable macromolecular conformations of a macromolecule, the method comprising: (i) isolating a population of macromolecules wherein the population of macromolecules comprises the at least two stable macromolecule conformations of the macromolecule, wherein the at least two stable macromolecule conformations of the macromolecule possess physical differences which allow the at least two stable macromolecule conformations to be separated from each other; (ii) separating the at least two stable macromolecule conformations from one another to provide at least two distinct homogeneous populations of purified macromolecules wherein each population is conformationally distinct from one another; (iii) reacting each conformationally distinct population of macromolecules with a covalent modifier which binds differentially to each of the stable macromolecule conformations, thereby providing covalently modified macromolecules which comprise newly created epitopes, wherein the newly created epitope comprises at least one bound covalent modifier; (iv) immunizing a laboratory animal; (v) monitoring the immune response to the immunogen preparation to determine a titer of antibodies to the newly created epitopes; (vi) collecting blood comprising the antibodies to the newly created epitope; (vii) preparing antisera; and (viii) fractionating the antisera to enrich for antibodies reactive to the newly created epitopes; thereby preparing antibodies that selectively bind to the newly created epitopes and thus are capable of discriminating between at least two stable macromolecular conformations of a macromolecule.