Highly Selective Separations in Biotechnology

1994
ISBN: 978-0-7514-0051-9
Verlag: Springer Netherlands

Buch, Englisch, 231 Seiten, HC runder Rücken kaschiert, Format (B × H): 160 mm x 241 mm, Gewicht: 1160 g

Success in meeting the challenge to produce the commercial products anticipated by the exploitation of biological processes depends upon provid­ ing effective separation protocols. Effectiveness can be measured in terms of selectivity, purity, resolution and validatory success. The major processing problems are associated with either the selective recovery of molecules which are present in low concentrations from complex mixtures or the selective removal of contaminants from the desired molecule. Central to the evolution of processes satisfying this demand are the regulatory requirements being imposed by governments on the purity of a product, especially in the health care market. Synthetic organic chemists are increasingly finding it advantageous to conduct one or more steps using either enzymic biotransformations where molecules with a single and consistent stereochemistry or chirality are required. The underlying princi­ ples behind the methods, techniques and processes currently being used and developed commercially rely upon the biospecific nature and properties of the desired molecule. When these factors are married to the more traditional techniques of precipitation, chromatography, liquid-liquid extraction and membrane processes, powerful tools emerge, allowing highly selective separations to be designed. The logical extension of these combinations is to apply genetic engineering techniques to influence the separations at a more fundamental and structural level by modifying the target protein at source, during its synthesis, to facilitate its separation in a given, selective manner, leading to the distinct possibility of producing 'designer' separation programmes.

Zielgruppe

Research

Autoren/Hrsg.

Street, G.

Fachgebiete

• Naturwissenschaften Biowissenschaften Mikrobiologie
• Technische Wissenschaften Verfahrenstechnik | Chemieingenieurwesen | Biotechnologie Verfahrenstechnik, Chemieingenieurwesen
• Technische Wissenschaften Verfahrenstechnik | Chemieingenieurwesen | Biotechnologie Biotechnologie Industrielle Biotechnologie
• Naturwissenschaften Biowissenschaften Tierkunde / Zoologie Tierethologie
• Naturwissenschaften Biowissenschaften Biochemie (nichtmedizinisch)

Inhaltsverzeichnis

1 Overview.- References.- 2 Affinity precipitation.- 2.1 Introduction.- 2.2 Various formats of affinity precipitation.- 2.3 Heterobifunctional ligands.- 2.4 Use of macro affinity ligands.- 2.5 Practical applications of affinity precipitation.- 2.6 Affinity precipitation by heterobifunctional ligands.- 2.7 Comparison of affinity precipitation with other affinity techniques.- 2.8 Conclusions.- References.- 3 Membrane-based affinity separation processes.- 3.1 Introduction.- 3.2 Chemical and physical feature of the membrane matrix.- 3.3 Purification protocol.- 3.4 Ligand coupling.- 3.5. Stability of active membranes.- 3.6 Reuse of ligand coupled membranes.- 3.7 Storage of ligand coupled membranes in buffer.- 3.8 Ligand leaching.- 3.9 Efficiency of ligand coupling.- 3.10 Effect of flow rates on affinity purification of IgG.- 3.11 Exhaustive purification of antibodies.- 3.12 Effect of sample recirculation on yields.- 3.13 Purification of IgG from ascites fluid and serum.- 3.14 Efficiency in product recovery.- 3.15 Applications.- 3.16 Scale-up.- 3.17 Discussion.- References.- 4 Affinity partitioning.- 4.1 Introduction.- 4.2 Two-phase systems.- 4.3 General ways of steering the partitioning.- 4.4 Affinity partitioning.- 4.5 Large-scale extractions.- 4.6 Use of two-phase systems in bioreactors.- 4.7 Non-protein partitioning.- 4.8 Alternative types of affinity ligands.- 4.9 Counter-current distribution.- 4.10 Conclusions.- References.- 5 The use of reverse micelles for the separation of proteins.- 5.1 Introduction.- 5.2 Description of reverse micelles.- 5.3 The reverse micellar extraction method.- 5.4 Protein distribution between an aqueous and a conjugated reverse micellar phase.- 5.5 Mass transfer of protein extraction.- 5.6 Process development.- 5.7 Examples of reverse micellarapplications for protein separation.- 5.8 Conclusions.- References.- 6 The chemistry and engineering of affinity chromatography.- 6.1 Introduction.- 6.2 The role of AC in protein purification.- 6.3 Affinity packings.- 6.4 Characterization of AC packings.- 6.5 Modelling and design of affinity chromatography columns.- 6.6 Notation.- References.- 7 Protein fusions as an aid to purification.- 7.1 Introduction.- 7.2 Choice of host organism.- 7.3 Induction of expression.- 7.4 Solubilisation of recombinant proteins.- 7.5 Vectors.- 7.6 Affinity purification.- 7.7 Thrombin cleavage of fusion proteins.- 7.8 General discussion.- References.- 8 Chiral separations.- 8.1 Introduction.- 8.2 Some stereochemical terms.- 8.3 Chirality and biological systems.- 8.4 Methods available for chiral separation.- 8.5 Conclusion.- References.- 9 Molecular imprinting—a versatile technique for the preparation of separation materials of predetermined selectivity.- 9.1 Introduction.- 9.2 Preparation of molecular imprints.- 9.3 Recognition in molecularly imprinted polymers.- 9.4 Application.- 9.5 Conclusions.- References.