Optimization Studies in Preparative Chromatography of Biomolecules

Optimization Studies in Preparative Chromatography of Biomolecules
Author: Sundar Ramanan
Publisher:
Total Pages: 254
Release: 2000
Genre: Biomolecules
ISBN:

Optimization of preparative nonlinear chromatography was carried out for the first time for a biomolecule mixture. Conventional wisdom on optimization, which roots from analytical chromatography, dictates optimizing resolution in an analytical column and obtaining similar separation in a large column for isolation. Such a method of optimization significantly under uses the capacity of the column and consumes large quantities of mobile phase. Hence, in preparative chromatography, the objective function is productivity, a measure of compromise between the amount of feed that can be loaded on to the column and time. Here, we report results from optimization studies carried out on a closely related binary peptide mixture on an analytical reversed-phase column. The goal is to optimize productivity under various chromatographic modes nonlinear isocratic elution, gradient elution, stepwise elution and displacement chromatography. In each mode, feed mixtures at highest possible concentration (limited by solubility), for increasing feed volumes was used. Productivity was monitored for increasing feed volumes, and loading was stopped as it went through a maximum. However, in some cases, solubility limitations from one of the feed components prevented further increase in loading. Even with this constraint, high productivities (5-10 g product/L stationary phase-h) were achieved. Separate experiments were carried out to measure the adsorption isotherms of these peptides over the range permitted by solubility. Separations under nonlinear chromatographic conditions were applied to isolate commercially significant two microcystins (microcystin LR and microcystin LA) from a cyanobacterial process waste. Milligram-level loading of microcystins was obtained on a solid-phase extraction cartridge packed with 0.5 g of C1 stationary phase. The separations were first carried out on an analytical column and then scaled-up to a preparative column. We also report simple and economical process to purify phycocyanins and allophycocyanins from a cyanobactenal process waste stream for two kinds of applications food colorant and biomedical marker. A detailed design for the large-scale production of biliproteins for both applications is also presented. Economic evaluation of the process resulted in comparable costs with the current market price for food-grade product and substantially lower cost for the biomedical grade product.

Scale-Up and Optimization in Preparative Chromatography

Scale-Up and Optimization in Preparative Chromatography
Author: Anurag Rathore
Publisher: CRC Press
Total Pages: 403
Release: 2002-09-26
Genre: Medical
ISBN: 1135564140

Presenting guidelines to predict and improve separation system performance, this book contains numerous case studies illustrating the practice of scale-up principles in process development. It offers solutions to limitations that occur in real-world purification schemes; methods to model, optimize, and characterize nonlinear separation processes; d

Scale-Up and Optimization in Preparative Chromatography

Scale-Up and Optimization in Preparative Chromatography
Author: Anurag Rathore
Publisher: CRC Press
Total Pages: 368
Release: 2002-09-26
Genre: Medical
ISBN: 9780203909287

Presenting guidelines to predict and improve separation system performance, this book contains numerous case studies illustrating the practice of scale-up principles in process development. It offers solutions to limitations that occur in real-world purification schemes; methods to model, optimize, and characterize nonlinear separation processes; d

Studies in Preparative Chromatography

Studies in Preparative Chromatography
Author: Ling Zhang
Publisher:
Total Pages: 268
Release: 2003
Genre: Countercurrent chromatography
ISBN:

Preparative chromatography is the most widely used purification method in the biotechnology and pharmaceutical industry. Here, one theoretical and one practical problem of preparative chromatography are investigated. The theoretical problem deals with the effect of experimental error on the precision of adsorption isotherm parameters estimated by numerical method. Numerical method has becoming popular in recent years as a quick alternative to the classical dynamic methods for adsorption isotherm determination. However, the fundamental problem of the numerical back-propagation of error has not been addressed in the literature. In this study, the variance in the final estimated parameters as a function of experimental error is quantified. The results indicate that it is theoretically possible to obtain sufficiently accurate parameters from multiple experimental replicates. The number of experimental replicates required to achieve isotherm parameters with less than 5% error is calculated explicitly for a variety of data sets. By using computer simulated experimental data, the effect of error in effluent concentrations and error in retention times are addressed separately. The practical problem is about the optimization of preparative purification of chlorophyll a by countercurrent chromatography (CCC). CCC offers several advantages over high performance liquid chromatography (HPLC) and is a fundamentally preparative separation tool. The process reported here is capable of producing several hundreds of milligrams of chlorophyll [alpha] with a high purity in a short time. The CCC separation is optimized for good production rate and recovery yield with respect to loading conditions.

Analytical and Preparative Separation Methods of Biomacromolecules

Analytical and Preparative Separation Methods of Biomacromolecules
Author: Hassan Y. Aboul-Enein
Publisher: CRC Press
Total Pages: 482
Release: 1999-05-26
Genre: Science
ISBN: 9780824719968

Reports up-to-date research developments on purifying and isolation large organic molecules. The text provides information on high-performance liquid chromatography and capillary electrophoresis (CE) as tools for analyzing biomacromolecules and developing new biochemical and medicinal compounds. It applies biochemical separation technology to the study of macromolecules such as proteins, polysaccharides, nucleic acids and more.

Preparative Chromatography

Preparative Chromatography
Author: H. Schmidt-Traub
Publisher: John Wiley & Sons
Total Pages: 648
Release: 2020-06-02
Genre: Science
ISBN: 3527344861

The third edition of this popular work is revised to include the latest developments in this fast-changing field. Its interdisciplinary approach elegantly combines the chemistry and engineering to explore the fundamentals and optimization processes involved.

Chromatography in Biotechnology

Chromatography in Biotechnology
Author: Csaba Horváth
Publisher:
Total Pages: 206
Release: 1993
Genre: Language Arts & Disciplines
ISBN:

The most up-to-date compilation of significant research on preparative liquid chromatography used for the separation of biomolecules and proteins. Presents recent advances in high-performance liquid chromatographic techniques for the isolation and purification of bioproducts in the laboratory and manufacturing plant. Discusses novel approaches to the preparative/process chromatography of complex carbohydrate and glycoconjugates. Also describes recent advances in column materials.

Preparative Chromatography for Separation of Proteins

Preparative Chromatography for Separation of Proteins
Author: Arne Staby
Publisher: John Wiley & Sons
Total Pages: 608
Release: 2017-02-02
Genre: Technology & Engineering
ISBN: 1119031176

Preparative Chromatography for Separation of Proteins addresses a wide range of modeling, techniques, strategies, and case studies of industrial separation of proteins and peptides. • Covers broad aspects of preparative chromatography with a unique combination of academic and industrial perspectives • Presents Combines modeling with compliantce useing of Quality-by-Design (QbD) approaches including modeling • Features a variety of chromatographic case studies not readily accessible to the general public • Represents an essential reference resource for academic, industrial, and pharmaceutical researchers

Preparative Chromatography

Preparative Chromatography
Author: H. Schmidt-Traub
Publisher: John Wiley & Sons
Total Pages: 714
Release: 2020-02-27
Genre: Science
ISBN: 352781633X

The third edition of this popular work is revised to include the latest developments in this fast-changing field. Its interdisciplinary approach elegantly combines the chemistry and engineering to explore the fundamentals and optimization processes involved.