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<strong>Column</strong>s for Bioanalytical ChromatographyBio-Monolith Protein A <strong>Column</strong> Rapidly Monitors IgG Fermentation Titers<strong>Column</strong>: Protein A5069-36395.2 x 4.95 mmMobile Phase: A: 1x PBS, pH 7.0B: 0.5 M acetic acid, pH 2.6Flow Rate: 1 mL/minGradient: 100% buffer A100% buffer B100% buffer A(0.5 min each step)Detector: UV at 280 nmInstrument: High pressure gradient<strong>HPLC</strong> system, <strong>Agilent</strong> 1200Relative absorbance (mAU)24019014090402 mL/min1mL/min_ 10Time (min)An IgG standard was measured at two flow rates, 1 mL/min and 2 mL/min, which correspond to 10 columnvolumes/min and 20 column volumes/min, respectively. The concentration of IgG in the sample can be reliablyobtained within minutes.16001400The calibration curve1200Peak area1000800600y = 728.55xR 2 = 0.99754002000C (mg/mL)The above calibration curve results from a 2-fold dilution series of human IgG concentrate. The initial IgGconcentration was 2 mg/mL. IgG peak areas were integrated and plotted. The linear range of the assay easilycovers the production ranges necessary to accommodate development and manufacturing cell cultures.130 Order online at www.agilent.com/chem/store
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