Electron microscopy-based semi-automated characterization of aggregation in monoclonal antibody products

Autor: Mohit Kumar, James Gomes, Ashutosh Pandey, Manidipa Banerjee, Kedar Khare, Rohit Bansal, Apoorv Pant, Anurag S. Rathore
Rok vydání: 2020
Předmět:
Connected component labelling
medicine.drug_class
lcsh:Biotechnology
HDX-MS
Hydrogen Deuterium Exchange Mass Spectroscopy
Biophysics
Protein aggregation
Monoclonal antibody
Biochemistry
Antibodies
law.invention
TV
Total Variation
Aggregation
03 medical and health sciences
0302 clinical medicine
MS
Mass Spectroscopy
Structural Biology
law
lcsh:TP248.13-248.65
UV
Ultra Violet
Electron microscopy
Genetics
medicine
DPBS
Dulbecco's phosphate-buffered saline
Cluster analysis
mAb
monoclonal Antibody
ComputingMethodologies_COMPUTERGRAPHICS
EM
Electron Microscopy
030304 developmental biology
0303 health sciences
FEG
field emission electron gun
TEM
Transmission Electron Microscopy
CD
Circular Dichroism
GUI
Graphical User Interface
Chemistry
Protein species
DLS
Dynamic Light Scattering
SEC-MALS
Size Exclusion Chromatography Multi Angle Light Scattering
Computer Science Applications
Characterization (materials science)
SEC
Size Exclusion Chromatography
Heterogeneous population
Critical parameter
030220 oncology & carcinogenesis
Heterogeneity
Electron microscope
Biological system
Research Article
Biotechnology
Zdroj: Computational and Structural Biotechnology Journal
Computational and Structural Biotechnology Journal, Vol 18, Iss, Pp 1458-1465 (2020)
ISSN: 2001-0370
Popis: Graphical abstract
Highlights • Size-based quantification of small heterogeneous proteins using electron microscopy. • Electron microscopy as an orthogonal tool for characterizing protein aggregates. • Quick assessment of small heterogeneous proteins via softEM, a GUI-based algorithm.
Aggregation is a critical parameter for protein-based therapeutics, due to its impact on the immunogenicity of the product. The traditional approach towards characterization of such products is to use a collection of orthogonal tools. However, the fact that none of these tools is able to completely classify the distribution and physical characteristics of aggregates, implies that there exists a need for additional analytical methods. We report one such method for characterization of heterogeneous population of proteins using transmission electron microscopy. The method involves semi-automated, size-based clustering of different protein species from micrographs. This method can be utilized for quantitative characterization of heterogeneous populations of antibody/protein aggregates from TEM images of proteins, and may also be applicable towards other instances of protein aggregation.
Databáze: OpenAIRE
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