Briefly, infiltrated 7 dpi whole-leaf samples were homogenized using a Matstone 6-in-one juice extractor (Matstone, PH), protein were extracted using PBS, pH 7.4 containing cOmplete™ ULTRA tablets mini, and EASYpack protease inhibitor cocktail (Roche, CH). The supernatant was clarified by a series of centrifugation and filtration steps. The bNAbs were then purified by protein A affinity chromatography using a HiTrap® Protein A High-Performance column (Cytiva, USA), coupled to a Äkta Avant 150 (Cytiva, USA), as per the manufacturer's instructions.
Kta avant 150
Manufactured by Cytiva
Sourced in United States
The Äkta Avant 150 is a versatile liquid chromatography system designed for preparative and process-scale purification of proteins, peptides, and other biomolecules. It is capable of operating at flow rates up to 150 mL/min and pressures up to 50 bar, making it suitable for a wide range of applications.
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2 protocols using kta avant 150
1
Purifying Antibodies from Infected Plant Leaves
2
Optimizing Elution Conditions for Affinity Chromatography
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Elution buffer optimization runs and frontal experiments for determination of the breakthrough curves were carried out in an ÄKTA™ avant 150 (Cytiva) equipped with a built-in fraction collector. Pre-packed OPUS® MiniChrom™ columns (Repligen, Waltham, MA, USA) with 0.2 mL of Capto AVB (Cytiva) were used. Prior to use, each column was regenerated with 50 CV of 10 mM of NaOH, followed by a cycle composed of 50 CV of equilibration buffer, 50 CV of elution buffer, and finally 50 CV of equilibration buffer.
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