Analytical control of enzyme-catalyzed peptide synthesis using capillary isotachophoresis

Stehle, Peter; Bahsitta, Hans-Peter; Fürst, Peter

doi:10.1016/s0021-9673(00)94681-8

Cited by 18 publications

(4 citation statements)

References 23 publications

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“…Great efforts were made to obtain reliable data concerning peptide purity. We established a novel free-flow electrophoretic technology (analytical isotachophoresis) enabling simultaneous determination of organic and inorganic impurities in the purified peptide material (Stehle et al 1986). The combined use of this highly specific method with reverse-phase HPLC techniques showed purity degrees Ͼ 99% for the synthetic peptides.…”

Section: Dipeptide Conceptmentioning

confidence: 99%

New Developments in Glutamine Delivery

Fürst

2001

The Journal of Nutrition

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Numerous studies demonstrate that free glutamine can be added to commercially available crystalline amino acid-based preparations before their administration. Instability during heat sterilization and prolonged storage and limited solubility (35 g/L at 20°C) hamper the use of free glutamine in the routine clinical setting. Indeed, there are many well-controlled and valuable trials with free glutamine, yet its use is restricted to clinical research. The obvious limitations of using free glutamine initiated an intensive search for alternative substrates. Synthetic glutamine dipeptides are stable under heat sterilization and highly soluble; these properties qualify the dipeptides as suitable constituents of nutritional preparations. Industrial production of these dipeptides at a reasonable price is an essential prerequisite for implications of dipeptide-containing solutions in clinical practice. Recent development of novel synthesis procedures allows increased capacity in industrial-scale production. Basic studies with synthetic glutamine-containing short-chain peptides provide convincing evidence that these new substrates are cleared rapidly from plasma after parenteral administration, without being accumulated in tissues and with negligible loss in urine. The presence of membrane-bound as well as tissue-free extracellular hydrolase activity facilitates a prompt and quantitative peptide hydrolysis, the liberated amino acids being available for protein synthesis and/or generation of energy. In the clinical setting, glutamine dipeptide nutrition beneficially influences outcome (nitrogen balance, immunity, gut integrity, hospital stay, morbidity and mortality). The provision of conditionally indispensable glutamine should be considered a necessary replacement of a deficiency rather than a supplementation. The beneficial effects observed with glutamine dipeptide nutrition should be seen simply as a correction of disadvantages produced by the inadequacy of conventional clinical nutrition. The availability of stable dipeptide preparations certainly facilitates, for the first time, adequate amino acid nutrition of critically ill, malnourished or stressed patients in the routine clinical setting and, thus, represents a new dimension in artificial nutrition.

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Section: Dipeptide Conceptmentioning

confidence: 99%

New Developments in Glutamine Delivery

Fürst

2001

The Journal of Nutrition

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“…Great efforts were made to ob tain reliable data concerning peptide purity. We established a novel free-flow electropho retic technology (analytical isotachophoresis) enabling simultaneous determination of or ganic and inorganic impurities in the purified peptide material [33], The combined use of this highly specific method with reverse-phase HPLC techniques showed purity degrees bet ter than 99% for the synthetic peptides.…”

Section: Syn Th E Sis and Characterization Of Parenteral Dipeptidesmentioning

confidence: 99%

Reappraisal of Indispensable Amino Acids

Fürst

Pogan²,

Hummel³

et al. 1997

Ann Nutr Metab

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“…Up to this point, ITP had mostly been limited to a preconcentration and separation technique. Then, in a number of publications led by researchers like Bocek 6,7 and Furst, 8,9 ITP applications were expanded to include the initiation and control of chemical reactions, particularly those involving enzymes and peptides. In those assays, ITP's high-resolving capability would be used to detect concentration changes in the enzyme-catalyzed mixture of reactants and products at several time points of the reaction.…”

Section: Introductionmentioning

confidence: 99%

Isotachophoresis applied to biomolecular reactions

Eid

Santiago

2018

Lab Chip

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This review discusses research developments and applications of isotachophoresis (ITP) to the initiation, control, and acceleration of chemical reactions, emphasizing reactions involving biomolecular reactants such as nucleic acids, proteins, and live cells. ITP is a versatile technique which requires no specific geometric design or material, and is compatible with a wide range of microfluidic and automated platforms. Though ITP has traditionally been used as a purification and separation technique, recent years have seen its emergence as a method to automate and speed up chemical reactions. ITP has been used to demonstrate up to 14 000-fold acceleration of nucleic acid assays, and has been used to enhance lateral flow and other immunoassays, and even whole bacterial cell detection assays. We here classify these studies into two categories: homogeneous (all reactants in solution) and heterogeneous (at least one reactant immobilized on a solid surface) assay configurations. For each category, we review and describe physical modeling and scaling of ITP-aided reaction assays, and elucidate key principles in ITP assay design. We summarize experimental advances, and identify common threads and approaches which researchers have used to optimize assay performance. Lastly, we propose unaddressed challenges and opportunities that could further improve these applications of ITP.

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Analytical control of enzyme-catalyzed peptide synthesis using capillary isotachophoresis

Cited by 18 publications

References 23 publications

New Developments in Glutamine Delivery

New Developments in Glutamine Delivery

Reappraisal of Indispensable Amino Acids

Isotachophoresis applied to biomolecular reactions

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