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• W1994522145 abstract "Locating optimal protein precipitation conditions for complex biological feed materials is problematic. This article describes the application of a series of high-throughput platforms for the rapid identification and selection of conditions for the precipitation of an IgG(4) monoclonal antibody (mAb) from a complex feedstock using only microliter quantities of material. The approach uses 96-microwell filter plates combined with high-throughput analytical methods and a method for well volume determination for product quantification. The low material, time and resource requirements facilitated the use of a full factorial Design of Experiments (DoE) for the rapid investigation into how critical parameters impact the IgG(4) precipitation. To aid the DoE, a set of preliminary range-finding studies were conducted first. Data collected through this approach describing Polyethylene Glycol (PEG) precipitation of the IgG(4) as a function of mAb concentration, precipitant concentration, and pH are presented. Response surface diagrams were used to explore interactions between parameters and to inform selection of the most favorable conditions for maximum yield and purification. PEG concentrations required for maximum yield and purity were dependant on the IgG(4) concentration; however, concentrations of 14 to 20% w/v, pH 6.5, gave optimal levels of yield and purity. Application of the high-throughput approach enabled 1,155 conditions to be examined with less than 1 g of material. The level of insights gained over such a short time frame is indicative of the power of microwell experimentation in allowing the rapid identification of appropriate processing conditions for key bioprocess operations." @default.
• W1994522145 created "2016-06-24" @default.
• W1994522145 creator A5000908646 @default.
• W1994522145 creator A5026913738 @default.
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• W1994522145 date "2009-12-11" @default.
• W1994522145 modified "2023-10-10" @default.
• W1994522145 title "High-throughput screening techniques for rapid PEG-based precipitation of IgG4 mAb from clarified cell culture supernatant" @default.
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• W1994522145 doi "https://doi.org/10.1002/btpr.357" @default.
• W1994522145 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/20014099" @default.
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