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 precious survey of exploratory tools regularly applied throughout the preclinical, scientific, and advertisement levels of improvement.
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Additional resources for Analytical techniques for biopharmaceutical development
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. 2 Protein transformations in reversed-phase chromatography for a two-state model. The native folded state can exist in either the mobile phase (Fm) or the stationary phase (Fs), as can the unfolded state (Um, Us).
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. However, note that high values of tG or low values of F will increase analysis time, may reduce recovery, and can induce conformational changes.
7). 8). For silica-based Copyright © 2005 by Marcel Dekker. 6 Effect of organic concentration on retention. W. R. R. Larsen, BioChromatography, 4: 19 . ) 45 Copyright © 2005 by Marcel Dekker. 7 Effect of high organic modifier concentration on peptide retention. Data obtained using an octadecyl column and 20 mM ammonium acetate:acetonitrile mobile phases. T. Wehr and L. Correia, LC at Work LC-121, Varian. 5 occurs by silanophilic interactions with the support matrix, with the roles of the aqueous and organic components of the mobile phase reversed to yield normal-phase chromatographic behavior.
Analytical techniques for biopharmaceutical development by Roberto Rodríguez-Díaz, Tim Wehr, Stephen F. Tuck