Direct affinity measurement of human IgG1 binding multimeric antigens

US12287335B2 · kind B2 · utility

Assignee

• Hoffmann-La Roche Inc. · US

Inventors

• Michael Molhoj · Munich, DE
• Christian Gassner · Penzberg, DE
• Joerg Moelleken · Penzberg, DE
• Manuel Endesfelder · Weßling, DE

Key dates

• Filing date: Oct 2, 2020
• Grant date: Apr 29, 2025
• Expiry date: Mar 10, 2043

Classification

• Technology area (CPC G): Physics
• CPC primary: G01N2333/96466
• WIPO field: Measurement
• WIPO sector: Instruments

Abstract

Herein is reported a method for determining the binding affinity of the binding sites of a bivalent full length antibody of the human IgG1 subclass to a homo-multimeric antigen comprising the steps of i) incubating a mixture comprising the antibody and a polypeptide that is derived from lysine-gingipain of Porphyromonas gingivalis at a pH of from pH 7.5 to pH 8.5, in the presence of a reducing agent, at a temperature of from 30° C. to 42° C., for time of from 10 min. to 240 min. to cleave the antibody into Fabs and Fc-region, and ii) determining the binding affinity of the Fabs of the antibody for its antigen using a surface plasmon resonance method by directly applying the incubated reaction mixture obtained in the previous step in the surface plasmon resonance method and therewith determining the binding affinity of the binding sites of the bivalent full length antibody of the human IgG1 subclass.