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<strong>Column</strong>s for Bioanalytical ChromatographyBio-Monolith DEAE <strong>Column</strong> Monitors Phage Production During Fermentation<strong>Column</strong>:Mobile Phase:Flow Rate:Gradient:Detector:Instrument:DEAE5069-36365.2 x 4.95 mmA: 125 mM Phosphate buffer,pH 7.0B: 125 mM Phosphate buffer+ 1M NaCl, pH 7.01 mL/min100% buffer A (2.5 min)0-100% buffer B (10 min)100% buffer A (2 min)UV at 280 nmHigh pressure gradient<strong>HPLC</strong> system, <strong>Agilent</strong> 1200Relative Absorbance (mAU)5045403530252015105Phages andother componentsTime 36 minTime 158 minTime 191 min% Buffer B13212010080604020% Gradient00 2 4 6 8 10 12Time (min)0As phage proliferation progresses, the genomic DNA (gDNA) concentration increases as the host cells are being lysed. In the late stages of fermentation gDNA begins todegrade into fragments. These gDNA fragments cannot be easily removed by purification media so, it is critical to stop the fermentation cycle prior to the degradation of thegenomic DNA. The chromatogram above represents three samples taken from the bioreactor at time 36 min, 158 min and 191 min. Peak 1 represents phage, media and hostcells, peak 2 the intact gDNA and peak 3 the fragmented gDNA.Order online at www.agilent.com/chem/store129
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