Hingst Elise J, Blech Michaela, Hinderberger Dariush, Garidel Patrick, Schwieger Christian
Institute of Chemistry, Physical Chemistry-Complex Self-Organizing Systems, Martin Luther University Halle-Wittenberg, Von-Danckelmann-Platz 4, 06120, Halle (Saale), Germany.
Boehringer Ingelheim Pharma GmbH & Co. KG. Innovation Unit, PDB-TIP, Birkendorfer Str. 65, 88397, Biberach (Riss), Germany.
Eur Biophys J. 2025 May 22. doi: 10.1007/s00249-025-01752-0.
Understanding the structure and self-organisation of monoclonal antibodies (mAbs) at the air-water interface is crucial for the stability and efficacy of protein drug formulations. This paper investigates the competitive adsorption of mAb and two amphiphilic polymers, poloxamer 188 (P188) and polysorbate 20 (PS20), commonly used to stabilise mAb formulations. Our objective was twofold: to ascertain whether the surfactants in question are capable of preventing mAb adsorption; and to determine whether it is possible to desorb mAb molecules from the air-water interface by surfactant addition. Langmuir film balance measurements and drop shape tensiometry were used to obtain surface pressure and surface tension data. Infrared Reflection-Absorption Spectroscopy (IRRAS) provided information on the surface composition, including the amount of adsorbed molecules. The state adopted by P188 is contingent upon its surface concentration, which determines the self-assembled phases it adopts. We show that the phase state of P188 has a considerable influence on mAb adsorption. The presence of P188 in the brush phase (≥ 0.3 mg/L) consistently inhibits mAb adsorption, but addition of P188 subsequent to the formation of the mAb film does not result in mAb desorption. However, addition of PS20 results in the desorption of freshly-formed interfacial mAb layers of up to two hours' age, whereas an aged mAb layer of 17 h was unable to be desorbed by PS20. Thus there is a time-dependent reorganisation of mAb at the air-water interface, increasing resistance to desorption, which we discuss in the context of potential intermolecular interactions within the interfacial film.
了解单克隆抗体（mAb）在空气-水界面的结构和自组装对于蛋白质药物制剂的稳定性和有效性至关重要。本文研究了mAb与两种两亲性聚合物泊洛沙姆188（P188）和聚山梨酯20（PS20）的竞争性吸附，这两种聚合物常用于稳定mAb制剂。我们的目标有两个：确定所讨论的表面活性剂是否能够防止mAb吸附；以及确定是否可以通过添加表面活性剂使mAb分子从空气-水界面解吸。使用Langmuir膜天平测量和液滴形状张力测定法来获取表面压力和表面张力数据。红外反射吸收光谱（IRRAS）提供了有关表面组成的信息，包括吸附分子的数量。P188所采用的状态取决于其表面浓度，表面浓度决定了它所采用的自组装相。我们表明，P188的相态对mAb吸附有相当大的影响。在刷状相（≥0.3 mg/L）中P188的存在始终抑制mAb吸附，但在mAb膜形成后添加P188不会导致mAb解吸。然而，添加PS20会导致新鲜形成的界面mAb层（长达两小时龄）解吸，而17小时龄的老化mAb层不能被PS20解吸。因此，mAb在空气-水界面存在时间依赖性的重组，增加了解吸阻力，我们将在界面膜内潜在的分子间相互作用的背景下对此进行讨论。
