Optimization of buffer conditions for the removal of endotoxins using Q-sepharose

So Hee Lee, Jung Soo Kim, Chan Wha Kim

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Anion-exchange chromatography using Q-sepharose was used to remove endotoxins. The optimized buffer conditions of pH, buffer concentration and NaCl concentration provided four log reductions in the endotoxin level (from 5 x 105 EU/ml to 10 EU/ml). In glycine-NaOH buffer (pH 8.0-10.5), the upper limit of the concentration of NaCl was 300 mM and that of glycine was 50 mM. For the Tris-HCl buffer (pH 7.5-9.0), the upper limit of the concentration of NaCl and buffer was 200 and 75 mM, respectively. In phosphate buffer (pH 6.0-8.0), the presence of phosphate significantly reduced the adsorption of endotoxins on Q-sepharose. The upper limit of the concentration of NaCl and phosphate in the phosphate buffer was 90 and 25 mM, respectively. The pH of the buffers showed very limited effects on the adsorption removal of endotoxins.

Original languageEnglish
Pages (from-to)1091-1098
Number of pages8
JournalProcess Biochemistry
Volume38
Issue number7
DOIs
Publication statusPublished - 2003 Feb 28

Fingerprint

Endotoxins
Sepharose
Buffers
Phosphates
Amino acids
Adsorption
Glycine
Chromatography
Ion exchange
Negative ions
Tromethamine
Anions

Keywords

  • Endotoxin removal
  • Phosphate buffer
  • Q-sepharose
  • Salt concentration

ASJC Scopus subject areas

  • Biochemistry

Cite this

Optimization of buffer conditions for the removal of endotoxins using Q-sepharose. / Lee, So Hee; Kim, Jung Soo; Kim, Chan Wha.

In: Process Biochemistry, Vol. 38, No. 7, 28.02.2003, p. 1091-1098.

Research output: Contribution to journalArticle

@article{39688890d20d4009b6ccd9295b9bdd95,
title = "Optimization of buffer conditions for the removal of endotoxins using Q-sepharose",
abstract = "Anion-exchange chromatography using Q-sepharose was used to remove endotoxins. The optimized buffer conditions of pH, buffer concentration and NaCl concentration provided four log reductions in the endotoxin level (from 5 x 105 EU/ml to 10 EU/ml). In glycine-NaOH buffer (pH 8.0-10.5), the upper limit of the concentration of NaCl was 300 mM and that of glycine was 50 mM. For the Tris-HCl buffer (pH 7.5-9.0), the upper limit of the concentration of NaCl and buffer was 200 and 75 mM, respectively. In phosphate buffer (pH 6.0-8.0), the presence of phosphate significantly reduced the adsorption of endotoxins on Q-sepharose. The upper limit of the concentration of NaCl and phosphate in the phosphate buffer was 90 and 25 mM, respectively. The pH of the buffers showed very limited effects on the adsorption removal of endotoxins.",
keywords = "Endotoxin removal, Phosphate buffer, Q-sepharose, Salt concentration",
author = "Lee, {So Hee} and Kim, {Jung Soo} and Kim, {Chan Wha}",
year = "2003",
month = "2",
day = "28",
doi = "10.1016/S0032-9592(02)00243-1",
language = "English",
volume = "38",
pages = "1091--1098",
journal = "Process Biochemistry",
issn = "1359-5113",
publisher = "Elsevier BV",
number = "7",

}

TY - JOUR

T1 - Optimization of buffer conditions for the removal of endotoxins using Q-sepharose

AU - Lee, So Hee

AU - Kim, Jung Soo

AU - Kim, Chan Wha

PY - 2003/2/28

Y1 - 2003/2/28

N2 - Anion-exchange chromatography using Q-sepharose was used to remove endotoxins. The optimized buffer conditions of pH, buffer concentration and NaCl concentration provided four log reductions in the endotoxin level (from 5 x 105 EU/ml to 10 EU/ml). In glycine-NaOH buffer (pH 8.0-10.5), the upper limit of the concentration of NaCl was 300 mM and that of glycine was 50 mM. For the Tris-HCl buffer (pH 7.5-9.0), the upper limit of the concentration of NaCl and buffer was 200 and 75 mM, respectively. In phosphate buffer (pH 6.0-8.0), the presence of phosphate significantly reduced the adsorption of endotoxins on Q-sepharose. The upper limit of the concentration of NaCl and phosphate in the phosphate buffer was 90 and 25 mM, respectively. The pH of the buffers showed very limited effects on the adsorption removal of endotoxins.

AB - Anion-exchange chromatography using Q-sepharose was used to remove endotoxins. The optimized buffer conditions of pH, buffer concentration and NaCl concentration provided four log reductions in the endotoxin level (from 5 x 105 EU/ml to 10 EU/ml). In glycine-NaOH buffer (pH 8.0-10.5), the upper limit of the concentration of NaCl was 300 mM and that of glycine was 50 mM. For the Tris-HCl buffer (pH 7.5-9.0), the upper limit of the concentration of NaCl and buffer was 200 and 75 mM, respectively. In phosphate buffer (pH 6.0-8.0), the presence of phosphate significantly reduced the adsorption of endotoxins on Q-sepharose. The upper limit of the concentration of NaCl and phosphate in the phosphate buffer was 90 and 25 mM, respectively. The pH of the buffers showed very limited effects on the adsorption removal of endotoxins.

KW - Endotoxin removal

KW - Phosphate buffer

KW - Q-sepharose

KW - Salt concentration

UR - http://www.scopus.com/inward/record.url?scp=0037681802&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037681802&partnerID=8YFLogxK

U2 - 10.1016/S0032-9592(02)00243-1

DO - 10.1016/S0032-9592(02)00243-1

M3 - Article

VL - 38

SP - 1091

EP - 1098

JO - Process Biochemistry

JF - Process Biochemistry

SN - 1359-5113

IS - 7

ER -