Subclass Effects on Self-Association and Viscosity of Monoclonal Antibodies at High Concentrations

被引:11
作者
Chowdhury, Amjad A. [1 ]
Manohar, Neha [1 ]
Witek, Marta A. [2 ]
Woldeyes, Mahlet A. [2 ]
Majumdar, Ranajoy [2 ]
Qian, Ken K. [2 ]
Kimball, William D. [1 ]
Xu, Shifeng [1 ]
Lanzaro, Alfredo [1 ]
Truskett, Thomas M. [1 ,3 ]
Johnston, Keith P. [1 ]
机构
[1] Univ Texas Austin, McKetta Dept Chem Engn, Austin, TX 78712 USA
[2] Eli Lilly & Co, Indianapolis, IN 46225 USA
[3] Univ Texas Austin, Dept Phys, Austin, TX 78712 USA
基金
美国国家卫生研究院; 美国国家科学基金会;
关键词
antibody subclasses; high concentration; viscosity; X-ray scattering; coarse-grained modeling; molecular descriptors; SMALL-ANGLE SCATTERING; X-RAY-SCATTERING; PROTEIN-PROTEIN INTERACTIONS; CLUSTER FORMATION; IGG1; AGGREGATION; BEHAVIOR; DISPERSIONS; STABILITY; DIFFUSION;
D O I
10.1021/acs.molpharmaceut.3c00023
中图分类号
R-3 [医学研究方法]; R3 [基础医学];
学科分类号
1001 ;
摘要
The effects of a subclass of monoclonal antibodies (mAbs)on protein-proteininteractions, formation of reversible oligomers (clusters), and viscosity(eta) are not well understood at high concentrations. Herein,we quantify a short-range anisotropic attraction between the complementarity-determiningregion (CDR) and CH3 domains (KCDR-CH3) for vedolizumabIgG1, IgG2, or IgG4 subclasses by fitting small-angle X-ray scattering(SAXS) structure factor S (eff)(q) data with an extensive library of 12-bead coarse-grained (CG) moleculardynamics simulations. The KCDR-CH3 bead attractionstrength was isolated from the strength of long-range electrostaticrepulsion for the full mAb, which was determined from the theoreticalnet charge and a scaling parameter psi to account for solventaccessibility and ion pairing. At low ionic strength (IS), the strongestshort-range attraction (KCDR-CH3) and consequentlythe largest clusters and highest eta were observed with IgG1,the subclass with the most positively charged CH3 domain. Furthermore,the trend in KCDR-CH3 with the subclass followedthe electrostatic interaction energy between the CDR and CH3 regionscalculated with the BioLuminate software using the 3D mAb structureand molecular interaction potentials. Whereas the equilibrium clustersize distributions and fractal dimensions were determined from fitsof SAXS with the MD simulations, the degree of cluster rigidity underflow was estimated from the experimental eta with a phenomenologicalmodel. For the systems with the largest clusters, especially IgG1,the inefficient packing of mAbs in the clusters played the largestrole in increasing eta, whereas for other systems, the relativecontribution from stress produced by the clusters was more significant.The ability to relate eta to short-range attraction from SAXSmeasurements at high concentrations and to theoretical characterizationof electrostatic patches on the 3D surface is not only of fundamentalinterest but also of practical value for mAb discovery, processing,formulation, and subcutaneous delivery.
引用
收藏
页码:2991 / 3008
页数:18
相关论文
共 84 条
  • [1] Masking of the Fc region in human IgG4 by constrained X-ray scattering modelling: implications for antibody function and therapy
    Abe, Yuki
    Gor, Jayesh
    Bracewell, Daniel G.
    Perkins, Stephen J.
    Dalby, Paul A.
    [J]. BIOCHEMICAL JOURNAL, 2010, 432 : 101 - 111
  • [2] Computational tool for the early screening of monoclonal antibodies for their viscosities
    Agrawal, Neeraj J.
    Helk, Bernhard
    Kumar, Sandeep
    Mody, Neil
    Sathish, Hasige A.
    Samra, Hardeep S.
    Buck, Patrick M.
    Li, Li
    Trout, Bernhardt L.
    [J]. MABS, 2016, 8 (01) : 43 - 48
  • [3] Modeling and mitigation of high-concentration antibody viscosity through structure-based computer-aided protein design
    Apgar, James R.
    Tam, Amy S. P.
    Sorm, Rhady
    Moesta, Sybille
    King, Amy C.
    Yang, Han
    Kelleher, Kerry
    Murphy, Denise
    D'Antona, Aaron M.
    Yan, Guoying
    Zhong, Xiaotian
    Rodriguez, Linette
    Ma, Weijun
    Ferguson, Darren E.
    Carven, Gregory J.
    Bennett, Eric M.
    Lin, Laura
    [J]. PLOS ONE, 2020, 15 (05):
  • [4] Suppression of protein interactions by arginine: A proposed mechanism of the arginine effects
    Arakawa, Tsutomu
    Ejima, Daisuke
    Tsumoto, Kouhei
    Obeyama, Noriyuki
    Tanaka, Yoshikazu
    Kita, Yoshiko
    Timasheff, Serge N.
    [J]. BIOPHYSICAL CHEMISTRY, 2007, 127 (1-2) : 1 - 8
  • [5] Charge-mediated Fab-Fc interactions in an IgG1 antibody induce reversible self-association, cluster formation, and elevated viscosity
    Arora, Jayant
    Hu, Yue
    Esfandiary, Reza
    Sathish, Hasige A.
    Bishop, Steven M.
    Joshi, Sangeeta B.
    Middaugh, C. Russell
    Volkin, David B.
    Weis, David D.
    [J]. MABS, 2016, 8 (08) : 1561 - 1574
  • [6] Predicting Antibody Developability Profiles Through Early Stage Discovery Screening
    Bailly, Marc
    Mieczkowski, Carl
    Juan, Veronica
    Metwally, Essam
    Tomazela, Daniela
    Baker, Jeanne
    Uchida, Makiko
    Kofman, Ester
    Raoufi, Fahimeh
    Motlagh, Soha
    Yu, Yao
    Park, Jihea
    Raghava, Smita
    Welsh, John
    Rauscher, Michael
    Raghunathan, Gopalan
    Hsieh, Mark
    Chen, Yi-Ling
    Nguyen, Hang Thu
    Nguyen, Nhung
    Cipriano, Dan
    Fayadat-Dilman, Laurence
    [J]. MABS, 2020, 12 (01)
  • [7] Blanco M. A., 2022, COMPUTATIONAL MODELS, P2044744
  • [8] HYDRODYNAMIC STRESS ON FRACTAL AGGREGATES OF SPHERES
    BOSSIS, G
    MEUNIER, A
    BRADY, JF
    [J]. JOURNAL OF CHEMICAL PHYSICS, 1991, 94 (07) : 5064 - 5070
  • [9] Highly Viscous Antibody Solutions Are a Consequence of Network Formation Caused by Domain-Domain Electrostatic Complementarities: Insights from Coarse-Grained Simulations
    Buck, Patrick M.
    Chaudhri, Anuj
    Kumar, Sandeep
    Singh, Satish K.
    [J]. MOLECULAR PHARMACEUTICS, 2015, 12 (01) : 127 - 139
  • [10] Predicting High-Concentration Interactions of Monoclonal Antibody Solutions: Comparison of Theoretical Approaches for Strongly Attractive Versus Repulsive Conditions
    Calero-Rubio, Cesar
    Saluja, Atul
    Sahin, Erinc
    Roberts, Christopher J.
    [J]. JOURNAL OF PHYSICAL CHEMISTRY B, 2019, 123 (27) : 5709 - 5720