Aggregation of biopharmaceuticals in human plasma and human serum

Abstract

Analytical methods based on light microscopy, 90° light-scattering and surface plasmon resonance (SPR) allowed the characterization of aggregation that can occur when antibodies are mixed with human plasma. Light microscopy showed that aggregates formed when human plasma was mixed with 5% dextrose solutions of Herceptin® (trastuzumab) or Avastin® (bevacizumab) but not Remicade® (infliximab). The aggregates in the plasma-Herceptin®-5% dextrose solution were globular, size range 0.5–9 μm, with a mean diameter of 4 μm. The aggregates in the plasma-Avastin®-5% dextrose samples had a mean size of 2 μm. No aggregation was observed when 0.9% NaCl solutions of Herceptin®, Avastin® and Remicade® were mixed with human plasma. 90° light-scattering measurements showed that aggregates were still present 2.5 h after mixing Herceptin® or Avastin® with 5% dextrose-plasma solution. A SPR method was utilized to qualitatively describe the extent of interactions of surface-bound antibodies with undiluted human serum. Increased binding was observed in the case of Erbitux® (cetuximab), whereas no binding was measured for Humira® (adalimumab). The binding of sera components to 13 monoclonal antibodies was measured and correlated with known serum binding properties of the antibodies. The data presented in this paper provide analytical methods to study the intrinsic and buffer-dependent aggregation tendencies of therapeutic proteins when mixed with human plasma and serum.

Considerations for the Design of Antibody-Based Therapeutics

“…Furthermore, mixing of formulated antibodies with serum can result in aggregation of antibodies with serum proteins in an antibody-and excipient-dependent manner. 329,330 Administration of certain antibody and formulation combinations could therefore result in aggregation-induced infusion reactions or loss of the effective drug. Differences in the number and size of aggregates have also been reported based on environment.…”

Surface plasmon resonance-based methodology for anti-adalimumab antibody identification and kinetic characterization

“…SPR-based optical biosensors detect in real-time mass changes on the chip surface, registering the interaction between the immobilized ligand and the analyte flowing in solution. SPR-based biosensors have been applied in a comparative study of affinities of ADA and other biologic drugs to the target TNF-α [26], or the aggregation tendencies of these therapeutic drugs when mixed with human plasma and serum [27]. However, a direct evaluation of anti-ADA antibodies and their affinity by means of SPR-based biosensors is to our knowledge not yet available.…”

Polymeric Micelles Loading Proteins through Concurrent Ion Complexation and pH‐Cleavable Covalent Bonding for In Vivo Delivery

“…Many therapeutic proteins suffer from poor pharmacokinetics due to aggregation in the bloodstream and rapid renal clearance . In this study, we used myoglobin, which is known to aggregate in blood and to be cleared by the kidneys, as the model protein for testing the ability of PEG‐p(Lys 20 ‐CDM 17 )‐based micelles to improve the proteins’ pharmacokinetics.…”