Investigating the Degradation Behaviors of a Therapeutic Monoclonal Antibody Associated with pH and Buffer Species.
This study aimed in understanding the degradation behaviors of an IgG 1 subtype therapeutic monoclonal antibody A (mAb-A) associated with pH and buffer species. The information obtained in this study can augment conventional, stability-based screening paradigms by providing the direction necessary for efficient experimental design. Differential scanning calorimetry (DSC) was used for studying conformational stability. Dynamic light scattering (DLS) was utilized to generate B22*, a modified second virial coefficient for the character of protein-protein interaction. Size-exclusion chromatography (SEC) and hydrophobic interaction chromatography (HIC) were employed to separate degradation products. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was used for determining the molecular size and liquid chromatography mass spectrometry (LC-MS) were used for identifying the sequence of the separated fragments. The results showed that both pH and buffer species played the roles in controlling the degradation behaviors of mAb-A, but the pH was more significant. In particular, pH 4.5 induced additional thermal transition peaks occurring at a low temperature compared with pH 6.5. A continual temperature-stress study illustrated that the additional thermal transition peaks related to the least stable structure and a greater fragmentation. Although mAb-A showed the comparable conformational structures and an identical amount of aggregates at time zero between the different types of buffer species at pH 6.5, the aggregation formation rate showed a buffer species-dependent discrepancy over a temperature-stress period. It was found that the levels of aggregations associated with the magnitudes of protein-protein interaction forces.
Drug Product Science and Technology, Bristol-Myers Squibb, One Squibb Drive, New Brunswick, New Jersey, 08903, USA. Songyan.zheng@bms.com.
Songyan
S
Zheng
Drug Product Science and Technology, Bristol-Myers Squibb, One Squibb Drive, New Brunswick, New Jersey, 08903, USA.
Difei
D
Qiu
Drug Product Science and Technology, Bristol-Myers Squibb, One Squibb Drive, New Brunswick, New Jersey, 08903, USA.
Monica
M
Adams
Drug Product Science and Technology, Bristol-Myers Squibb, One Squibb Drive, New Brunswick, New Jersey, 08903, USA.
Jinjiang
J
Li
Drug Product Science and Technology, Bristol-Myers Squibb, One Squibb Drive, New Brunswick, New Jersey, 08903, USA.
Rao V
RV
Mantri
Drug Product Science and Technology, Bristol-Myers Squibb, One Squibb Drive, New Brunswick, New Jersey, 08903, USA.
Rajesh
R
Gandhi
D000911:Antibodies, Monoclonal / Q000737:chemistry*; D002021:Buffers; D002152:Calorimetry, Differential Scanning / Q000379:methods; D002850:Chromatography, Gel / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006863:Hydrogen-Ion Concentration; D057927:Hydrophobic and Hydrophilic Interactions; D007074:Immunoglobulin G / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods; D011487:Protein Conformation; D013696:Temperature
D000911:Antibodies, Monoclonal; D002021:Buffers; D007074:Immunoglobulin G
aggregation; conformational stability; fragmentation; monoclonal antibody (mAb); protein stability; thermal stability