Verification of the final anion exchange chromatography in the r-hgh manufacturing process

Byeong Jo Min, Seong Woo Kang, Yoon Seok Song, Jong Ho Lee, Seung Heon Lee, Chulhwan Park, Seung Wook Kim, Chan Wha Kim

Research output: Contribution to journalArticle

Abstract

In this study, final anion exchange chromatography in the recombinant human growth hormone (r-hGH) manufacturing process was validated using a validation protocol that was consistent with both policy and standard operation procedure (SOP). Two buffer solutions used in chromatography were first validated and were found to satisfy pre-established acceptance criteria as follows: pH: 8.2, endotoxin: < 0.6 EU/mL, bioburden test: negative. Final anion exchange chromatography was conducted using a DEAE Sepharose FF Resin and eluted with a linear gradient of 30 to 110 mM NaCl in 50 mM Tris-HCl buffer at a flow rate of 15 L/h. Three consecutive batches of hGH solutions were generated via anion exchange chromatography, which was performed within pre-established operating parameters determined through in-process control. When all three batches were assessed by the pre-established sampling plan and tested for quality control, this purification process was shown to satisfy pre-established acceptance criteria; endotoxin: ≤0.5 EU/mg, ECP: ≤ 1.4 ppm, IEF: same removal distance, hGH content by Native-PAGE: 100%, purity by HPLC: ≥ 99%, yield by UV scanning: 87 to 89%, hGH monomer protein content by HPLC: 99%. Therefore, the final anion chromatography process was successfully validated in this study, and this method consistently yielded hGH solutions that satisfied pre-established criteria for subsequent processing.

Original languageEnglish
Pages (from-to)488-496
Number of pages9
JournalBiotechnology and Bioprocess Engineering
Volume15
Issue number3
DOIs
Publication statusPublished - 2010 Jun 1

Fingerprint

Chromatography
Anions
Ion exchange
Negative ions
Endotoxins
Buffers
High Pressure Liquid Chromatography
Native Polyacrylamide Gel Electrophoresis
Tromethamine
Human Growth Hormone
Hormones
Quality Control
Sepharose
Growth Hormone
Process control
Purification
Quality control
Resins
Monomers
Flow rate

Keywords

  • Acceptance criteria
  • Anion exchange chromatography
  • Recombinant human growth hormone (r-hGH)
  • Validation

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology
  • Biomedical Engineering
  • Bioengineering

Cite this

Verification of the final anion exchange chromatography in the r-hgh manufacturing process. / Min, Byeong Jo; Kang, Seong Woo; Song, Yoon Seok; Lee, Jong Ho; Lee, Seung Heon; Park, Chulhwan; Kim, Seung Wook; Kim, Chan Wha.

In: Biotechnology and Bioprocess Engineering, Vol. 15, No. 3, 01.06.2010, p. 488-496.

Research output: Contribution to journalArticle

Min, Byeong Jo ; Kang, Seong Woo ; Song, Yoon Seok ; Lee, Jong Ho ; Lee, Seung Heon ; Park, Chulhwan ; Kim, Seung Wook ; Kim, Chan Wha. / Verification of the final anion exchange chromatography in the r-hgh manufacturing process. In: Biotechnology and Bioprocess Engineering. 2010 ; Vol. 15, No. 3. pp. 488-496.
@article{f7e743c14c7445a3b3f38e9335767003,
title = "Verification of the final anion exchange chromatography in the r-hgh manufacturing process",
abstract = "In this study, final anion exchange chromatography in the recombinant human growth hormone (r-hGH) manufacturing process was validated using a validation protocol that was consistent with both policy and standard operation procedure (SOP). Two buffer solutions used in chromatography were first validated and were found to satisfy pre-established acceptance criteria as follows: pH: 8.2, endotoxin: < 0.6 EU/mL, bioburden test: negative. Final anion exchange chromatography was conducted using a DEAE Sepharose FF Resin and eluted with a linear gradient of 30 to 110 mM NaCl in 50 mM Tris-HCl buffer at a flow rate of 15 L/h. Three consecutive batches of hGH solutions were generated via anion exchange chromatography, which was performed within pre-established operating parameters determined through in-process control. When all three batches were assessed by the pre-established sampling plan and tested for quality control, this purification process was shown to satisfy pre-established acceptance criteria; endotoxin: ≤0.5 EU/mg, ECP: ≤ 1.4 ppm, IEF: same removal distance, hGH content by Native-PAGE: 100{\%}, purity by HPLC: ≥ 99{\%}, yield by UV scanning: 87 to 89{\%}, hGH monomer protein content by HPLC: 99{\%}. Therefore, the final anion chromatography process was successfully validated in this study, and this method consistently yielded hGH solutions that satisfied pre-established criteria for subsequent processing.",
keywords = "Acceptance criteria, Anion exchange chromatography, Recombinant human growth hormone (r-hGH), Validation",
author = "Min, {Byeong Jo} and Kang, {Seong Woo} and Song, {Yoon Seok} and Lee, {Jong Ho} and Lee, {Seung Heon} and Chulhwan Park and Kim, {Seung Wook} and Kim, {Chan Wha}",
year = "2010",
month = "6",
day = "1",
doi = "10.1007/s12257-009-3053-9",
language = "English",
volume = "15",
pages = "488--496",
journal = "Biotechnology and Bioprocess Engineering",
issn = "1226-8372",
publisher = "Korean Society for Biotechnology and Bioengineering",
number = "3",

}

TY - JOUR

T1 - Verification of the final anion exchange chromatography in the r-hgh manufacturing process

AU - Min, Byeong Jo

AU - Kang, Seong Woo

AU - Song, Yoon Seok

AU - Lee, Jong Ho

AU - Lee, Seung Heon

AU - Park, Chulhwan

AU - Kim, Seung Wook

AU - Kim, Chan Wha

PY - 2010/6/1

Y1 - 2010/6/1

N2 - In this study, final anion exchange chromatography in the recombinant human growth hormone (r-hGH) manufacturing process was validated using a validation protocol that was consistent with both policy and standard operation procedure (SOP). Two buffer solutions used in chromatography were first validated and were found to satisfy pre-established acceptance criteria as follows: pH: 8.2, endotoxin: < 0.6 EU/mL, bioburden test: negative. Final anion exchange chromatography was conducted using a DEAE Sepharose FF Resin and eluted with a linear gradient of 30 to 110 mM NaCl in 50 mM Tris-HCl buffer at a flow rate of 15 L/h. Three consecutive batches of hGH solutions were generated via anion exchange chromatography, which was performed within pre-established operating parameters determined through in-process control. When all three batches were assessed by the pre-established sampling plan and tested for quality control, this purification process was shown to satisfy pre-established acceptance criteria; endotoxin: ≤0.5 EU/mg, ECP: ≤ 1.4 ppm, IEF: same removal distance, hGH content by Native-PAGE: 100%, purity by HPLC: ≥ 99%, yield by UV scanning: 87 to 89%, hGH monomer protein content by HPLC: 99%. Therefore, the final anion chromatography process was successfully validated in this study, and this method consistently yielded hGH solutions that satisfied pre-established criteria for subsequent processing.

AB - In this study, final anion exchange chromatography in the recombinant human growth hormone (r-hGH) manufacturing process was validated using a validation protocol that was consistent with both policy and standard operation procedure (SOP). Two buffer solutions used in chromatography were first validated and were found to satisfy pre-established acceptance criteria as follows: pH: 8.2, endotoxin: < 0.6 EU/mL, bioburden test: negative. Final anion exchange chromatography was conducted using a DEAE Sepharose FF Resin and eluted with a linear gradient of 30 to 110 mM NaCl in 50 mM Tris-HCl buffer at a flow rate of 15 L/h. Three consecutive batches of hGH solutions were generated via anion exchange chromatography, which was performed within pre-established operating parameters determined through in-process control. When all three batches were assessed by the pre-established sampling plan and tested for quality control, this purification process was shown to satisfy pre-established acceptance criteria; endotoxin: ≤0.5 EU/mg, ECP: ≤ 1.4 ppm, IEF: same removal distance, hGH content by Native-PAGE: 100%, purity by HPLC: ≥ 99%, yield by UV scanning: 87 to 89%, hGH monomer protein content by HPLC: 99%. Therefore, the final anion chromatography process was successfully validated in this study, and this method consistently yielded hGH solutions that satisfied pre-established criteria for subsequent processing.

KW - Acceptance criteria

KW - Anion exchange chromatography

KW - Recombinant human growth hormone (r-hGH)

KW - Validation

UR - http://www.scopus.com/inward/record.url?scp=77956220276&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77956220276&partnerID=8YFLogxK

U2 - 10.1007/s12257-009-3053-9

DO - 10.1007/s12257-009-3053-9

M3 - Article

VL - 15

SP - 488

EP - 496

JO - Biotechnology and Bioprocess Engineering

JF - Biotechnology and Bioprocess Engineering

SN - 1226-8372

IS - 3

ER -