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By Roberto Rodríguez-Díaz, Tim Wehr, Stephen F. Tuck

This reference assists scientists within the choice and alertness of analytical concepts for biopharmaceutical identity, formula, and assessment-providing a invaluable survey of exploratory equipment regularly applied in the course of the preclinical, medical, and advertisement stages of improvement.

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Extra resources for Analytical techniques for biopharmaceutical development

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6. , Methods in Enzymology, 182: 587 (1990). Copyright © 2005 by Marcel Dekker. 4 Use of Reversed-Phase Liquid Chromatography in Biopharmaceutical Development Tim Wehr INTRODUCTION Reversed-phase liquid chromatography (RPLC) is an essential tool for analytical and preparative separations of biopharmaceuticals. Its ubiquitous application in the discovery and development of protein-based drugs arises from its high resolving power, the robustness of the separation technique, and its success in handling a wide range of separation problems.

R. Snyder, Anal. , 58: 280 [1986]. ) 3. Select a solvent range (∆Φ) for a scouting gradient. 4. Using the values of S, k*, and ∆Φ of in combination with Vm and F, estimate tG from Equation 6. 44 . 5. Run the scouting gradient, then narrow the gradient range so that the earliest peak elutes after t0 + dwell time (tD) and the last peak elutes near the final solvent concentration. The gradient steepness should be maintained in the narrowed gradient. Copyright © 2005 by Marcel Dekker. 50 Wehr 6. Fine tune tG, F, and L for best resolution while keeping k* constant.

S. Hodges, J. , 359: 519 [1986]. ) phase, and this process of solvent exclusion is the driving force for protein binding to the hydrocarbonaceous phase. 2). , in the millisecond domain), the observed peak represents the average of the rapidly interconverting species. If the interconversion rate is slow (seconds to minutes), varying contact areas of the different conformational states may be manifested by broadened peaks or chromatographic resolution of discrete peaks. Copyright © 2005 by Marcel Dekker.

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