The effect of pH, hydrogen peroxide and temperature on the stability of human monoclonal antibody.

The stability of human monoclonal antibody (C23), which is being developed as a passive immunotherapeutic agent against human cytomegalovirus, was investigated. C23 (about 2 mg ml-1) was incubated under sterile conditions for 14 days in buffers with different pH values (ranging from 4-10), in hydrogen peroxide solutions with different concentrations (0.01% or 0.1%), and in saline at 8 degrees C or 37 degrees C. Samples were collected on days 0, 3, 7 and 14, and various physicochemical or biological methods were used to determine the changes in in C23. These methods included turbidity (absorbance at 408 nm and transmittance at 580 nm), pH, size-exclusion high performance liquid chromatography (HPLC), hydroxyapatite HPLC, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, isoelectric focusing, matrix-assisted laser desorption ionization time-of-flight mass spectrometry and virus neutralization assay. Using these methods, the possible degradation processes of C23 were initially characterized. Deamidation, oxidation, fragmentation, covalent cross-links and aggregation were observed as major degradation routes. These results gave useful information for the manufacturing process and quality control of C23.