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KMID : 1094720100150030488
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Biotechnology and Bioprocess Engineering
2010 Volume.15 No. 3 p.488 ~ p.496
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Verification of the Final Anion Exchange Chromatography in the r-hGH Manufacturing Process
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Min Byeong-Jo
Kang Seong-Woo
Song Yoon-Seok
Lee Jong-Ho
Lee Seung-Heon
Park Chul-Hwan
Kim Seung-Wook
Kim Chan-Wha
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Abstract
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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.
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KEYWORD
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validation, recombinant human growth hormone (r-hGH), acceptance criteria, anion exchange chromatography
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