Automated recycling free fluid isotachophoresis

Sloan, Jeffrey E.; Thormann, Wolfgang; Twitty, G. E.; Bier, M.

doi:10.1016/s0021-9673(01)82061-6

Cited by 15 publications

(2 citation statements)

References 19 publications

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“…Another way to overcome the problems of long separation times along with short transit times was the development of recycling isotachophoresis (R-ITP) [18]. As in CF-ITP, fluid flows rapidly through a narrow channel and the effluent from each channel is reinjected into the electrophoresis chamber through the corresponding port of input.…”

Section: Introductionmentioning

confidence: 99%

Automated free‐solution isotachophoresis: Instrumentation and fractionation of human serum proteins

Böttcher¹,

Möllers²,

Lackner³

et al. 1998

Electrophoresis

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An automated free-solution isotachophoresis system (FS-ITP) for preparative fractionation of biopolymers is described, operated in a batch mode. The dimension of the separation chamber allows an up to 1200-fold higher sample load compared to separation in capillaries of 180 microm inner diameter as used in analytical capillary isotachophoresis (C-ITP). The preparative capacity of the system is within the milligram range. The method is fully compatible with analytical C-ITP, which is essential for preparative-scale isotachophoresis with regard to optimization of electrolyte systems and the search for suitable spacers. As a model application the fractionation of human serum proteins is reported. The collected fractions were analyzed by C-ITP and agarose gel electrophoresis.

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Section: Introductionmentioning

confidence: 99%

Automated free‐solution isotachophoresis: Instrumentation and fractionation of human serum proteins

Böttcher¹,

Möllers²,

Lackner³

et al. 1998

Electrophoresis

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“…Preparative instruments based on continuous, and recycling fluid flow, as well as on rotating fluid are commercially available [l-51. In recycling operation, processing fluid leaving the separation cell through the output ports is recycled with a peristaltic pump and reinjected into the electrophoresis chamber through appropriate inlet ports. Instrumentation and protein purification based on recycling zone electrophoresis (RZE) [6], recycling isotachophoresis (RITP) [7] and recycling isoelectric focusing (RIEF) [S], have been described. Most recycling devices are modularly built, and thus can be easily adapted for operation in different electrophoretic modes.…”

Section: Introductionmentioning

confidence: 99%

Isolation of human serum transferrin by free‐fluid recycling electrophoresis in simple buffers

Caslavska¹,

Thormann

1994

Electrophoresis

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The isolation of human serum transferrin (Tf) using recycling isotachophoresis (RITP) and recycling isoelectric focusing (RIEF) with simple buffers is described. Serum fractionation, the first step in the protocol for Tf purification, is shown to be easily performed either by RITP of filtered serum using low molecular mass spacers or by RIEF of dialyzed serum employing a binary mixture of a well-defined buffer pair covering the pH range between 5.2 and 6.2, called RIEF-OptiFocus. For polishing, Tf-containing fractions are reprocessed by RITP or RIEF-OptiFocus. Other RIEF approaches based on the use of single amino acids at high concentration and ternary amino acid mixtures are shown to constitute less effective methods for Tf isolation. With a four-step protocol, comprising in turn RITP, RIEF-OptiFocus, ultrafiltration and again RITP, "single-band" purity (as assessed by two-dimensional gel electrophoresis) is obtained. Omission of the first step (RITP) and direct processing of dialyzed serum by RIEF-OptiFocus, ultrafiltration and RITP, is shown to provide remarkable results as well.

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Isotachophoretic zone formation of serum albumin in different free fluid electrophoresis instruments

et al. 1990

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The isotachophoretic behavior of a model protein, serum albumin, was examined (i) by computer simulation, (ii) by capillary isotachophoresis in HPE 100 and Tachophor 2127, (iii) by continuous flow isotachophoresis in Elphor VaP 22 and the BIO-STREAM Separator and (iv) by recycling isotachophoresis in an apparatus of our own design. Variations in monitored zone shapes can be explained by differences in engineering aspects and fluid stabilization principles of the instruments.

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Automated recycling free fluid isotachophoresis

Cited by 15 publications

References 19 publications

Automated free‐solution isotachophoresis: Instrumentation and fractionation of human serum proteins

Automated free‐solution isotachophoresis: Instrumentation and fractionation of human serum proteins

Isolation of human serum transferrin by free‐fluid recycling electrophoresis in simple buffers

Isotachophoretic zone formation of serum albumin in different free fluid electrophoresis instruments

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