Probing protein aggregation at buried interfaces: distinguishing between adsorbed protein monomers, dimers, and a monomer–dimer mixture <i>in situ</i>

Lu, Tieyi; Prathamesh, Datar M.; Xin, Yue; Marsh, E. Neil G.; Chen, Zhan

doi:10.1039/d1sc04300e

Cited by 13 publications

(20 citation statements)

References 85 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In this study, to find the most probable antibody orientation, the calculated SFG spectra and the reconstructed resonant spectra from experimental data are normalized in intensity (with the highest signal in the spectrum) to make the spectral feature comparison for ssp or ppp spectrum. The details of the Hamiltonian matching process have been published before and will not be repeated here. , The most likely antibody orientation can be determined by an overall heat map which combines the heat maps for the ssp spectrum, ppp spectrum, and ssp/ppp signal strength ratio.…”

Section: Resultsmentioning

confidence: 99%

“…The details of the Hamiltonian matching process have been published before and will not be repeated here. 11,37 The most likely antibody orientation can be determined by an overall heat map which combines the heat maps for the ssp spectrum, ppp spectrum, and ssp/ppp signal strength ratio.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

“…SFG is a surface-sensitive characterization method that allows us to probe the structural information of molecules on surfaces or at interfaces, − which can be used to study the protein structures ,,− including antibodies on silicone oil surfaces . The detail of SFG theory was published before − and is discussed briefly in the Supporting Information, Section S1. In this study, a femtosecond SFG system was applied to study antibody–silicone oil interactions.…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Probing Molecular Interactions of Antibody Drugs, Silicone Oil, and Surfactant at Buried Interfaces In Situ

Qiu

et al. 2022

Anal. Chem.

View full text Add to dashboard Cite

Antibody drugs have been rapidly developed to cure many diseases including COVID-19 infection. Silicone oil is commonly used as a lubricant coating material for devices used in the pharmaceutical industry to store and administer antibody drug formulations. However, the interaction between silicone oil and antibody molecules could lead to the adsorption, denaturation, and aggregation of antibody molecules, impacting the efficacy of antibody drugs. Here, we studied the molecular interactions between antibodies and silicone oil in situ in real time. The effect of the surfactant on such interactions was also investigated. Specifically, the adsorption dynamics of a bispecific antibody (BsAb) onto a silicone oil surface without and with different concentrations of the surfactant PS80 in antibody solutions were monitored. Also the possible lowest effective PS80 concentrations that can prevent the adsorption of BsAb as well as a monoclonal antibody (mAb) onto silicone oil were measured. It was found that different concentrations of PS80 are required for preventing the adsorption of different antibodies. Both BsAB and mAB denature on silicone oil without a surfactant. However, for a low surfactant concentration in the solution, although the surfactant could not completely prevent the antibody from adsorption, it could maintain the native structures of adsorbed BsAb and mAb antibodies on silicone oil. This is important knowledge, showing that to prevent antibody aggregation on silicone oil it is not necessary to add surfactant to a concentration high enough to completely minimize protein adsorption.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: ■ Results and Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Probing Molecular Interactions of Antibody Drugs, Silicone Oil, and Surfactant at Buried Interfaces In Situ

Qiu

et al. 2022

Anal. Chem.

View full text Add to dashboard Cite

show abstract

“…The stepping stone of the current research is based on the characterization of the two species that form the two ends of the phase‐separation spectrum—the monomer and the dimer—of the AXH domain which is predicted to contribute to the aggregation pathway of ATXN1 in addition to the PolyQ chain 36 . Rather than considering the higher order aggregates, we here focus on the dimeric species considering it as the inception point or the first glimpse of the self‐association process, that is, the logic behind the foundation is that the dimeric species originates as the product of the 1st order self‐association process 76–77 . Additionally, the advantage of studying a dimeric species comes from the fact that it's the simplest form of species that can appear upon self‐association and thus makes it the least complex structure that one can deal with.…”

Section: Discussionmentioning

confidence: 99%

Governing dynamics and preferential binding of the AXH domain influence the aggregation pathway of Ataxin‐1

Tammara

Das

2022

Proteins

View full text Add to dashboard Cite

The present state of understanding the mechanism of Spinocerebellar Ataxia‐1, a fatal neurodegenerative disease linked to the protein Ataxin‐1 (ATXN1), is baffled by a set of self‐contradictory, and hence, inconclusive observations. This fallacy poses a bottleneck to the effective designing of curable drugs as the field is currently missing the specific druggable site. To understand the fundamentals of pathogenesis, we tried to decipher the intricacies of the extremely complicated landscape by targeting the relevant species that supposedly dictate the structure–function paradigm. The atomic‐level description and characterization of the dynamism of the systems reveal the existence of structural polymorphism in all the leading stakeholders of the overall system. The very existence of conformational heterogeneity in every species creates numerous possible combinations of favorable interactions because of the variability in segmental cross‐talks and hence claims its role in the choice of routes between functional activity and dysfunctional disease‐causing aggregation. Despite this emergent configurational diversity, there is a common mode of operative intermolecular forces that dictates the extent of stability of all the multimeric complexes due to the localized population of a specific type of residue. The present research proposes a dynamic switch mechanism between aggregability and functional activity, based on the logical interpretation of the estimated variables, which is practically dictated by the effective concentration of the interacting species involved in the cell.

show abstract

“…The SFG spectra were collected using ppp (p-polarized sum frequency beam, p-polarized visible beam, and p-polarized infrared beam) polarization combination. The basic theory of the SFG has been extensively described in a lot of studies, − and a brief introduction of the SFG and relative equations (eqs S1–S9) used for the calculation can be found in the Supporting Information.…”

Section: Methodsmentioning

confidence: 99%

Enhanced Sum Frequency Generation for Monolayers on Au Relative to Silica: Local Field Factors and SPR Effect

Han

et al. 2022

Langmuir

View full text Add to dashboard Cite

Using metals as signal magnified substrates, surface plasmon-enhanced sum frequency generation (SFG) vibrational spectroscopy is a promising technique to probe weak molecular-level signals at surfaces and interfaces. In this study, the vibrational signals of the n-alkane monolayer on the gold (Au) and silica substrates are investigated using the broadband femtosecond SFG. The enhancement factors are discovered to be up to ∼1076 and ∼31 for the methyl symmetric and asymmetric stretching (ss and as) modes of the monolayer, respectively. By systematically analyzing the second-order nonlinear susceptibility tensor components (χ ijk s), the Fresnel coefficients (F ijk s), and the surface plasmon resonance (SPR) effect, we find that the interplay between F ijk and χ ijk terms and the SPR effect dominate the SFG signal enhancement. Our study reveals that the relative contributions of different influencing factors (i.e., Fresnel coefficients and SPR) to the SFG signal enhancement provide an approach to interpreting enhanced SFG vibrational signals detected from probe molecules on distinct substrates and may finally guide the design of the experimental methodology to improve the detection sensitivity and signal-to-noise ratio.

show abstract

Probing protein aggregation at buried interfaces: distinguishing between adsorbed protein monomers, dimers, and a monomer–dimer mixture in situ

Abstract: Protein adsorption on surfaces greatly impacts many applications such as biomedical materials, anti-biofouling coatings, bio-separation membranes, biosensors, and antibody protein drugs etc. For example, protein drug adsorption on widely used...

Cited by 13 publications

References 85 publications

Probing Molecular Interactions of Antibody Drugs, Silicone Oil, and Surfactant at Buried Interfaces In Situ

Probing Molecular Interactions of Antibody Drugs, Silicone Oil, and Surfactant at Buried Interfaces In Situ

Governing dynamics and preferential binding of the AXH domain influence the aggregation pathway of Ataxin‐1

Enhanced Sum Frequency Generation for Monolayers on Au Relative to Silica: Local Field Factors and SPR Effect

Contact Info

Product

Resources

About