Nature and consequences of protein–protein interactions in high protein concentration solutions

Saluja, Atul; Kalonia, Devendra S.

doi:10.1016/j.ijpharm.2008.03.041

Cited by 188 publications

(178 citation statements)

References 127 publications

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“…[5][6][7] The RSA of mAbs presents various pharmaceutical challenges, including the formation of protein aggregation precursors that can lead to the irreversible formation of oligomers. 8,9 In addition, the RSA of mAbs gives rise to a network of associated higher-order species that can affect the viscoelastic properties of the solution, 10 resulting in increased viscosity, [5][6][7]11 solution turbidity, 12 and, under certain conditions, even phase transitions. 13 The increase in solution viscosity also imposes manufacturing challenges including high back-pressure and clogging of membranes, 2 as well as elevated levels of shear stress during pumping.…”

Section: Introductionmentioning

confidence: 99%

Hydrogen exchange mass spectrometry reveals protein interfaces and distant dynamic coupling effects during the reversible self-association of an IgG1 monoclonal antibody

Arora

Hickey

Majumdar

et al. 2015

mAbs

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Volkin (2015) Hydrogen exchange mass spectrometry reveals protein interfaces and distant dynamic coupling effects during the reversible self-association of an IgG1 monoclonal antibody, mAbs, 7:3, 525-539, DOI: 10.1080DOI: 10. /19420862.2015 To link to this article: https://doi.org/10. 1080/19420862.2015 There is a need for new analytical approaches to better characterize the nature of the concentration-dependent, reversible self-association (RSA) of monoclonal antibodies (mAbs) directly, and with high resolution, when these proteins are formulated as highly concentrated solutions. In the work reported here, hydrogen exchange mass spectrometry (HX-MS) was used to define the concentration-dependent RSA interface, and to characterize the effects of association on the backbone dynamics of an IgG1 mAb (mAb-C). Dynamic light scattering, chemical cross-linking, and solution viscosity measurements were used to determine conditions that caused the RSA of mAb-C. A novel HX-MS experimental approach was then applied to directly monitor differences in local flexibility of mAb-C due to RSA at different protein concentrations in deuterated buffers. First, a stable formulation containing lyoprotectants that permitted freeze-drying of mAb-C at both 5 and 60 mg/mL was identified. Upon reconstitution with RSA-promoting deuterated solutions, the low vs. high protein concentration samples displayed different levels of solution viscosity (i.e., approx. 1 to 75 mPa.s). The reconstituted mAb-C samples were then analyzed by HX-MS. Two specific sequences covering complementarity-determining regions CDR2H and CDR2L (in the variable heavy and light chains, respectively) showed significant protection against deuterium uptake (i.e., decreased hydrogen exchange). These results define the major protein-protein interfaces associated with the concentration-dependent RSA of mAb-C. Surprisingly, certain peptide segments in the V H domain, the constant domain (C H 2), and the hinge region (C H 1-C H 2 interface) concomitantly showed significant increases in local flexibility at high vs. low protein concentrations. These results indicate the presence of longer-range, distant dynamic coupling effects within mAb-C occurring upon RSA.

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Section: Introductionmentioning

confidence: 99%

Hydrogen exchange mass spectrometry reveals protein interfaces and distant dynamic coupling effects during the reversible self-association of an IgG1 monoclonal antibody

Arora

Hickey

Majumdar

et al. 2015

mAbs

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“…29 Rather than relying on cumulative effects on retention time in the chromatographic methods to detect these weak interactions, sensitive BLI technology 22 allows for direct monitoring of antibody self-binding. CSI-BLI has unparalleled throughput compared with CIC or SIC in that a plate of 96 antibodies can be tested for self-binding within 2 h, rather than a few days.…”

Section: High Throughput Detection Of Antibody Self-interaction By Bimentioning

confidence: 99%

High throughput detection of antibody self-interaction by bio-layer interferometry

Sun

Reid

Liu

et al. 2013

mAbs

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“…Nonspecific interactions contribute not only to the stochastic diffusion of macromolecules (Saluja and Kalonia 2008;de Nooijer et al 2009;Johansson et al 2014;Staneva and Frenkel 2015;Luchinat and Banci 2017), but also to the formation of localized structures such as organelles and supramolecular assemblies (Lécuyer et al 2007;Rudner and Losick 2010;Coquel et al 2013;Wei et al 2016;Luo et al 2016). Hence, they determine the macroscopic properties of the cytoplasm both in physiological conditions and under stress (Parry et al 2014;Joyner et al 2016;Munder et al 2016).…”

Section: Introductionmentioning

confidence: 99%

Molecular simulations of cellular processes

Trovato

Fumagalli

2017

Biophys Rev

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It is, nowadays, possible to simulate biological processes in conditions that mimic the different cellular compartments. Several groups have performed these calculations using molecular models that vary in performance and accuracy. In many cases, the atomistic degrees of freedom have been eliminated, sacrificing both structural complexity and chemical specificity to be able to explore slow processes. In this review, we will discuss the insights gained from computer simulations on macromolecule diffusion, nuclear body formation, and processes involving the genetic material inside cell-mimicking spaces. We will also discuss the challenges to generate new models suitable for the simulations of biological processes on a cell scale and for cellcycle-long times, including non-equilibrium events such as the co-translational folding, misfolding, and aggregation of proteins. A prominent role will be played by the wise choice of the structural simplifications and, simultaneously, of a relatively complex energetic description. These challenging tasks will rely on the integration of experimental and computational methods, achieved through the application of efficient algorithms.

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Nature and consequences of protein–protein interactions in high protein concentration solutions

Cited by 188 publications

References 127 publications

Hydrogen exchange mass spectrometry reveals protein interfaces and distant dynamic coupling effects during the reversible self-association of an IgG1 monoclonal antibody

Hydrogen exchange mass spectrometry reveals protein interfaces and distant dynamic coupling effects during the reversible self-association of an IgG1 monoclonal antibody

High throughput detection of antibody self-interaction by bio-layer interferometry

Molecular simulations of cellular processes

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