Kanthe Ankit, Ilott Andrew, Krause Mary, Zheng Songyan, Li Jinjiang, Bu Wei, Bera Mrinal K, Lin Binhua, Maldarelli Charles, Tu Raymond S
Department of Chemical Engineering, The City College of New York, New York, NY 10031, USA.
Drug Product Development, Bristol Myers Squibb, New Brunswick, NJ 08901, USA.
Sci Adv. 2021 Aug 27;7(35). doi: 10.1126/sciadv.abg2873. Print 2021 Aug.
The interaction of monoclonal antibodies (mAbs) with air/water interfaces plays a crucial role in their overall stability in solution. We aim to understand this behavior using pendant bubble measurements to track the dynamic tension reduction and x-ray reflectivity to obtain the electron density profiles (EDPs) at the surface. Native immunoglobulin G mAb is a rigid molecule with a flat, "Y" shape, and simulated EDPs are obtained by rotating a homology construct at the surface. Comparing simulations with experimental EDPs, we obtain surface orientation probability maps showing mAbs transition from flat-on Y-shape configurations to side-on or end-on configurations with increasing concentration. The modeling also shows the presence of β sheets at the surface. Overall, the experiments and the homology modeling elucidate the orientational phase space during different stages of adsorption of mAbs at the air/water interface. These finding will help define new strategies for the manufacture and storage of antibody-based therapeutics.
单克隆抗体（mAb）与空气/水界面的相互作用在其在溶液中的整体稳定性中起着关键作用。我们旨在通过悬垂气泡测量来跟踪动态张力降低，并利用X射线反射率来获取表面的电子密度分布（EDP），以了解这种行为。天然免疫球蛋白G单克隆抗体是一种具有扁平“Y”形的刚性分子，通过在表面旋转同源构建体来获得模拟的EDP。将模拟结果与实验EDP进行比较，我们得到了表面取向概率图，显示随着浓度增加，单克隆抗体从平躺的Y形构型转变为侧躺或端对端构型。建模还表明表面存在β折叠。总体而言，实验和同源建模阐明了单克隆抗体在空气/水界面吸附不同阶段的取向相空间。这些发现将有助于确定基于抗体的治疗药物制造和储存的新策略。
