Fate of antibody and polysorbate particles in a human serum model

Human Serum Albumin Mitigates Formation of Fatty Acid Particles in Polysorbate-Containing Solutions

“…Recent studies have evaluated the fate of polysorbate-derived particles in mAb formulations in the presence of human plasma, serum, or infusion solutions. [18][19][20][21] The findings presented here build upon these previous studies by providing a reasonable mechanism leading to the observed phenomena. The mechanism of particle dissolution in infusion solutions perhaps differs from that in serum, plasma, or in the in vitro conditions described here; despite these differences, a similar end point (reversal of fatty acid particles) was achieved.…”

Combined Presence of Ferrous Ions and Hydrogen Peroxide in Normal Saline and In Vitro Models Induces Enhanced Aggregation of Therapeutic IgG due to Hydroxyl Radicals

“…Normal saline is isotonic and consists of 0.9% NaCl . However, once the sample containing mAb is administered in the body, it is exposed to various salts, pH of 7.4, and different small and large molecules. ,− Hence, to determine the aggregation of mAb samples with FeSO 4 ·7H 2 O and H 2 O 2 after administration in the body, stability was investigated in extracellular saline and more complex in vitro models.…”

“…The SEC chromatogram of HSF (also shown in Figure S1a) (and FBSF) further showed a set of peaks after 37 min. It could be due to the presence of small peptides and amino acids, and these peaks were not in the analysis range of mAb and aggregates. , The serum filtrate (FBSF and HSF) was used to analyze stability of mAb. The low %HMW of mAb spiked in HSF and FBSF containing Fe 2+ and H 2 O 2 compared to samples in saline and extracellular saline might be due to the difference in the electrolyte composition, pH, and presence of molecules of physiological origin such as amino acids, peptides, carbohydrates, lipids, and electrolytes, along with other small molecules including thiols in macromolecule free serum.…”

“…Monoclonal antibodies (mAb) are an important class of protein therapeutics that undergo stability-related issues in the form of aggregation and fragmentation during various stages of their life cycle. − These size-based heterogeneities are believed to negatively impact activity and shelf life of the product , and potentially result in adverse immune responses. ,, Hence, monitoring and control of aggregates are of utmost importance. , The aggregates could be soluble or insoluble. While soluble aggregates include oligomeric species (<100 nm size), the insoluble aggregates consist of particles of subvisible (0.1–100 μm) and visible (>100 μm) size range .…”

“…Furthermore, the stability of mAb in physiological fluids after its administration into the human body might be different from the stability in buffers and solutions used during formulation, manufacturing, and administration . Hence, various buffer systems and in vitro models such as serum and macromolecule free fraction of serum have been used to determine the stability of mAb under conditions that are physiologically relevant. , The difference in the stability of mAb in the presence of both Fe 2+ and H 2 O 2 in fluids, such as artificially prepared extracellular saline vs macromolecule free fraction of serum has not yet been explored. Moreover, increased levels of Fe 2+ and H 2 O 2 are also found in the human body during various diseases such as myocardial infarction, neurodegenerative diseases, uremia, and diabetes mellitus, − ,− and the stability of mAb could be altered in patients with elevated levels of Fe 2+ and H 2 O 2 compared to patients who do not have any diseases concerned with elevated levels of Fe 2+ and H 2 O 2 .…”