Electrostatics Drive Oligomerization and Aggregation of Human Interferon Alpha-2a
Pohl, Christin LU ; Polimeni, Marco LU ; Indrakumar, Sowmya ; Streicher, Werner ; Peters, Günther H.J. ; Nørgaard, Allan ; Lund, Mikael LU
and Harris, Pernille
(2021)
In Journal of Physical Chemistry B
125(50).
p.13657-13669
- Abstract
Aggregation is a common phenomenon in the field of protein therapeutics and can lead to function loss or immunogenic patient responses. Two strategies are currently used to reduce aggregation: (1) finding a suitable formulation, which is labor-intensive and requires large protein quantities, or (2) engineering the protein, which requires extensive knowledge about the protein aggregation pathway. We present a biophysical characterization of the oligomerization and aggregation processes by Interferon alpha-2a (IFNα-2a), a protein drug with antiviral and immunomodulatory properties. This study combines experimental high throughput screening with detailed investigations by small-angle X-ray scattering and analytical ultracentrifugation.... (More)
Aggregation is a common phenomenon in the field of protein therapeutics and can lead to function loss or immunogenic patient responses. Two strategies are currently used to reduce aggregation: (1) finding a suitable formulation, which is labor-intensive and requires large protein quantities, or (2) engineering the protein, which requires extensive knowledge about the protein aggregation pathway. We present a biophysical characterization of the oligomerization and aggregation processes by Interferon alpha-2a (IFNα-2a), a protein drug with antiviral and immunomodulatory properties. This study combines experimental high throughput screening with detailed investigations by small-angle X-ray scattering and analytical ultracentrifugation. Metropolis Monte Carlo simulations are used to gain insight into the underlying intermolecular interactions. IFNα-2a forms soluble oligomers that are controlled by a fast pH and concentration-dependent equilibrium. Close to the isoelectric point of 6, IFNα-2a forms insoluble aggregates which can be prevented by adding salt. We show that monomer attraction is driven mainly by molecular anisotropic dipole–dipole interactions that increase with increasing pH. Repulsion is due to monopole–monopole interactions and depends on the charge of IFNα-2a. The study highlights how combining multiple methods helps to systematically dissect the molecular mechanisms driving oligomer formation and to design ultimately efficient strategies for preventing detrimental protein aggregation.
(Less)
- author
- Pohl, Christin
LU
; Polimeni, Marco
LU
; Indrakumar, Sowmya
; Streicher, Werner
; Peters, Günther H.J.
; Nørgaard, Allan
; Lund, Mikael
LU
and Harris, Pernille
- organization
- publishing date
- 2021-12-23
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of Physical Chemistry B
- volume
- 125
- issue
- 50
- pages
- 13 pages
- publisher
- The American Chemical Society (ACS)
- external identifiers
-
- pmid:34898211
- scopus:85121709623
- ISSN
- 1520-6106
- DOI
- 10.1021/acs.jpcb.1c07090
- language
- English
- LU publication?
- yes
- additional info
- Publisher Copyright: © 2021 The Authors. Published by American Chemical Society.
- id
- 46632d26-4a70-4920-817f-3c57a1a245cf
- date added to LUP
- 2022-01-20 10:01:08
- date last changed
- 2024-04-26 02:21:23
@article{46632d26-4a70-4920-817f-3c57a1a245cf,
abstract = {{<p>Aggregation is a common phenomenon in the field of protein therapeutics and can lead to function loss or immunogenic patient responses. Two strategies are currently used to reduce aggregation: (1) finding a suitable formulation, which is labor-intensive and requires large protein quantities, or (2) engineering the protein, which requires extensive knowledge about the protein aggregation pathway. We present a biophysical characterization of the oligomerization and aggregation processes by Interferon alpha-2a (IFNα-2a), a protein drug with antiviral and immunomodulatory properties. This study combines experimental high throughput screening with detailed investigations by small-angle X-ray scattering and analytical ultracentrifugation. Metropolis Monte Carlo simulations are used to gain insight into the underlying intermolecular interactions. IFNα-2a forms soluble oligomers that are controlled by a fast pH and concentration-dependent equilibrium. Close to the isoelectric point of 6, IFNα-2a forms insoluble aggregates which can be prevented by adding salt. We show that monomer attraction is driven mainly by molecular anisotropic dipole–dipole interactions that increase with increasing pH. Repulsion is due to monopole–monopole interactions and depends on the charge of IFNα-2a. The study highlights how combining multiple methods helps to systematically dissect the molecular mechanisms driving oligomer formation and to design ultimately efficient strategies for preventing detrimental protein aggregation.</p>}},
author = {{Pohl, Christin and Polimeni, Marco and Indrakumar, Sowmya and Streicher, Werner and Peters, Günther H.J. and Nørgaard, Allan and Lund, Mikael and Harris, Pernille}},
issn = {{1520-6106}},
language = {{eng}},
month = {{12}},
number = {{50}},
pages = {{13657--13669}},
publisher = {{The American Chemical Society (ACS)}},
series = {{Journal of Physical Chemistry B}},
title = {{Electrostatics Drive Oligomerization and Aggregation of Human Interferon Alpha-2a}},
url = {{http://dx.doi.org/10.1021/acs.jpcb.1c07090}},
doi = {{10.1021/acs.jpcb.1c07090}},
volume = {{125}},
year = {{2021}},
}