scholarly journals Parallel chromatography and in situ scattering to interrogate competing protein aggregation pathways

2018 ◽  
Vol 27 (7) ◽  
pp. 1325-1333 ◽  
Author(s):  
Diana Gomes ◽  
Rebecca K. Kalman ◽  
Rebecca K. Pagels ◽  
Miguel A. Rodrigues ◽  
Christopher J. Roberts
Keyword(s):  
Protein Aggregation ◽  
In Situ Scattering

scholarly journals Inducing protein aggregation by extensional flow

2017 ◽  
Vol 114 (18) ◽  
pp. 4673-4678 ◽  
Author(s):  
John Dobson ◽  
Amit Kumar ◽  
Leon F. Willis ◽  
Roman Tuma ◽  
Daniel R. Higazi ◽  
...  
Keyword(s):  
Flow Field ◽  
Protein Aggregation ◽  
Large Scale ◽  
Extensional Flow ◽  
Cysteine Residue ◽  
Hydrodynamic Flow ◽  
Extrinsic Factors ◽  
Aggregation Propensity ◽  
Industrial Manufacture

Relative to other extrinsic factors, the effects of hydrodynamic flow fields on protein stability and conformation remain poorly understood. Flow-induced protein remodeling and/or aggregation is observed both in Nature and during the large-scale industrial manufacture of proteins. Despite its ubiquity, the relationships between the type and magnitude of hydrodynamic flow, a protein’s structure and stability, and the resultant aggregation propensity are unclear. Here, we assess the effects of a defined and quantified flow field dominated by extensional flow on the aggregation of BSA, β2-microglobulin (β2m), granulocyte colony stimulating factor (G-CSF), and three monoclonal antibodies (mAbs). We show that the device induces protein aggregation after exposure to an extensional flow field for 0.36–1.8 ms, at concentrations as low as 0.5 mg mL−1. In addition, we reveal that the extent of aggregation depends on the applied strain rate and the concentration, structural scaffold, and sequence of the protein. Finally we demonstrate the in situ labeling of a buried cysteine residue in BSA during extensional stress. Together, these data indicate that an extensional flow readily unfolds thermodynamically and kinetically stable proteins, exposing previously sequestered sequences whose aggregation propensity determines the probability and extent of aggregation.

scholarly journals Probing Protein Aggregation at Buried Interfaces: Distinguishing Adsorbed Protein Monomer, Dimer, and Monomer-Dimer Mixture in Situ

2022 ◽  
Author(s):  
Tieyi Lu ◽  
Wen Guo ◽  
Datar M. Prathamesh ◽  
Yue Xin ◽  
E. Neil G. Marsh ◽  
...  
Keyword(s):  
Protein Aggregation ◽  
Protein Adsorption ◽  
Protein Drug ◽  
Protein Drugs ◽  
Biomedical Materials ◽  
Drug Adsorption ◽  
Separation Membranes ◽  
Buried Interfaces ◽  
Adsorbed Protein

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...

Corrosion Product Identification by Micro-Raman and Mossbauer Spectroscopy

1998 ◽  
Vol 4 (S2) ◽  
pp. 506-507
Author(s):  
F. Adar ◽  
B. Lenain ◽  
D. C. Cook ◽  
S. J. Oh
Keyword(s):  
Mössbauer Spectroscopy ◽  
Mossbauer Spectroscopy ◽  
Spot Size ◽  
Corrosion Products ◽  
Raman Spectrometry ◽  
Mößbauer Spectroscopy ◽  
Metallographic Cross Section ◽  
Steel Coupon ◽  
In Situ Scattering

Micro-Raman spectrometry and Mossbauer spectroscopy have been used to identify the corrosion products on a steel coupon exposed in an industrial environment for 16 years. The Raman analysis was performed on a polished metallographic cross-section in order to map the oxides across the thickness of the coating. The spectra were recorded using a LabRam Micro-Raman spectrograph incorporating a 17 mW HeNe laser (attenuated to 1 mW to prevent oxide transformation), focused to 1 μm spot size, and 1800 g/mm grating. The confocal line-scan imaging enabled 100 spectra to be recorded in one scan at 0.5 um intervals across the thickness of the coating. The Mossbauer analysis was performed using in-situ scattering Mossbauer spectroscopy on the attached corrosion coating and transmission Mossbauer spectroscopy at 300K and 77K on the removed coating, to measure the fraction of each oxide present. Micro-Raman spectrometry showed that the corrosion products had formed in distinct layers as shown in Figure 1.

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