The stabilization of a human IgM monoclonal antibody with poly(vinylpyrrolidone)

Pharm Res. 1994 May;11(5):624-32. doi: 10.1023/a:1018903624373.

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)

MeSH terms

  • Antibodies, Monoclonal / chemistry*
  • Calorimetry, Differential Scanning
  • Chromatography, High Pressure Liquid
  • Drug Stability
  • Enzyme-Linked Immunosorbent Assay
  • Excipients
  • Hot Temperature
  • Immunoglobulin M / chemistry*
  • Light
  • Molecular Weight
  • Povidone / chemistry*
  • Scattering, Radiation

Substances

  • Antibodies, Monoclonal
  • Excipients
  • Immunoglobulin M
  • Povidone