title: The stabilization of a human IgM monoclonal antibody with poly(vinylpyrrolidone).
abstract: An IgM anti-group B Streptococcus monoclonal antibody (4B9) was found to undergo irreversible heat-induced aggregation at 50 degrees C. A variety of excipients was tested for their ability to inhibit antibody aggregation. The amount of 4B9 aggregation, which was determined by analysis on a size-exclusion HPLC, was significantly reduced in the presence of low concentrations [between 0.1 and 1.0% (w/v)] of poly(vinylpyrrolidone) (PVP) molecules ranging in molecular weight from 10 to 40 kDa. When the PVP concentration was greater than 1.0%, antibody aggregation was enhanced, and with the highest molecular weight PVP, antibody precipitation occurred. HPLC was used to show that more PVP was associated with the 4B9 at 50 degrees C than at 25 degrees C. Differential scanning calorimetry revealed that PVP concentrations greater than 2.0% decreased the antibody thermal transition temperature. Enzyme-linked immunosorbent assays were used to assess the effects of PVP on the antigen binding capacity of 4B9 and on 4B9 quantitation. At 4 degrees C, PVP solutions of up to 5.0% had no effect on either 4B9 quantitation or antigen binding. At 50 degrees C, however, less 4B9 was detected in the 5.0% PVP solution. The heat stabilization of the 4B9 antibody by low concentrations of PVP can be explained by a weak binding of PVP to the native protein. The PVP may sterically interfere with protein-protein interactions, thus reducing aggregation. Higher concentrations of PVP lead to protein aggregation and precipitation, probably by a volume-exclusion mechanism.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Bristol-Myers Squibb, Pharmaceutical Research Institute, Seattle, Washington 98121.
authors.forename: W R
authors.initials: WR
authors.lastname: Gombotz
authors.forename: S C
authors.initials: SC
authors.lastname: Pankey
authors.forename: D
authors.initials: D
authors.lastname: Phan
authors.forename: R
authors.initials: R
authors.lastname: Drager
authors.forename: K
authors.initials: K
authors.lastname: Donaldson
authors.forename: K P
authors.initials: KP
authors.lastname: Antonsen
authors.forename: A S
authors.initials: AS
authors.lastname: Hoffman
authors.forename: H V
authors.initials: HV
authors.lastname: Raff
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry*; D002152:Calorimetry, Differential Scanning; D002851:Chromatography, High Pressure Liquid; D004355:Drug Stability; D004797:Enzyme-Linked Immunosorbent Assay; D005079:Excipients; D006358:Hot Temperature; D007075:Immunoglobulin M / Q000737:chemistry*; D008027:Light; D008970:Molecular Weight; D011205:Povidone / Q000737:chemistry*; D012542:Scattering, Radiation
chemical_list: D000911:Antibodies, Monoclonal; D005079:Excipients; D007075:Immunoglobulin M; D011205:Povidone