Light-scattering detection within the difficult size range of protein particle measurement using flow cytometry. 2018

Zhishang Hu, and Chen Ye, and Wei Mi, and Yun Zhao, and Can Quan, and William W Li, and Hongmei Li, and Haiying Hang
Chemical Metrology & Analytical Science Division (NRCCRM), National Institute of Metrology, Beijing, China. lihm@nim.ac.cn.

The phenomenon of protein aggregation is a prominent challenge that impacts biopharmaceutical development at every stage. It may have a number of deleterious effects on protein drugs, including the loss of efficacy, induction of immunogenicity, altered pharmacokinetics and reduced shelf life. At present, multiple methods are available for counting and sizing particles over a broad range of sizes. However, there remains a conundrum in the measurement of particles in the submicrometer range, from 100 nm to 2 μm. In this study, the capability of our new laboratory built FCM system to detect model polystyrene (PS) and silica (SiO2) submicrometer microspheres was evaluated and benchmarked against flow field-flow fractionation (FFF). The FCM system showed its advantages on sensitivity, selectivity, reproducibility and speed. The laboratory-built FCM system can readily analyze model PS and SiO2 microspheres down to 200 nm, covering much of the difficult range from 100 nm to 2 μm. Our data also showed that this machine was able to monitor the distribution of antibody aggregates ranged between 200 nm and 10 μm, suggesting its usability for characterizing protein aggregation in future.

UI MeSH Term Description Entries
D010316 Particle Size Relating to the size of solids. Particle Sizes,Size, Particle,Sizes, Particle
D011137 Polystyrenes Polymerized forms of styrene used as a biocompatible material, especially in dentistry. They are thermoplastic and are used as insulators, for injection molding and casting, as sheets, plates, rods, rigid forms and beads. Polystyrol,Polystyrene,Polystyrols
D005434 Flow Cytometry Technique using an instrument system for making, processing, and displaying one or more measurements on individual cells obtained from a cell suspension. Cells are usually stained with one or more fluorescent dyes specific to cell components of interest, e.g., DNA, and fluorescence of each cell is measured as it rapidly transverses the excitation beam (laser or mercury arc lamp). Fluorescence provides a quantitative measure of various biochemical and biophysical properties of the cell, as well as a basis for cell sorting. Other measurable optical parameters include light absorption and light scattering, the latter being applicable to the measurement of cell size, shape, density, granularity, and stain uptake. Cytofluorometry, Flow,Cytometry, Flow,Flow Microfluorimetry,Fluorescence-Activated Cell Sorting,Microfluorometry, Flow,Cell Sorting, Fluorescence-Activated,Cell Sortings, Fluorescence-Activated,Cytofluorometries, Flow,Cytometries, Flow,Flow Cytofluorometries,Flow Cytofluorometry,Flow Cytometries,Flow Microfluorometries,Flow Microfluorometry,Fluorescence Activated Cell Sorting,Fluorescence-Activated Cell Sortings,Microfluorimetry, Flow,Microfluorometries, Flow,Sorting, Fluorescence-Activated Cell,Sortings, Fluorescence-Activated Cell
D000906 Antibodies Immunoglobulin molecules having a specific amino acid sequence by virtue of which they interact only with the ANTIGEN (or a very similar shape) that induced their synthesis in cells of the lymphoid series (especially PLASMA CELLS).
D012822 Silicon Dioxide Transparent, tasteless crystals found in nature as agate, amethyst, chalcedony, cristobalite, flint, sand, QUARTZ, and tridymite. The compound is insoluble in water or acids except hydrofluoric acid. Silica,Aerosil,Aerosil 380,Cristobalite,Quso G-32,Quso G32,Tridymite,380, Aerosil,Dioxide, Silicon,G32, Quso,Quso G 32
D059629 Signal-To-Noise Ratio The comparison of the quantity of meaningful data to the irrelevant or incorrect data. Ratio, Signal-To-Noise,Ratios, Signal-To-Noise,Signal To Noise Ratio,Signal-To-Noise Ratios
D031521 Fractionation, Field Flow Separation of molecules and particles by a simultaneous action of carrier liquid flow and focusing field forces (electrical, sedimentation, or thermal), without a stationary phase. Field Flow Fractionation,Sedimentation Field Flow Fractionation,SdFFF
D066329 Protein Aggregates Any mixture of secondary, tertiary, or quaternary protein molecules which appear as clumps in or outside the cell. Protein Aggregate,Aggregate, Protein,Aggregates, Protein

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