Verification of the Final Anion Exchange Chromatography in the r-hGH Manufacturing Process

• Published in: Biotechnology and bioprocess engineering 2010, 15(3), , pp.488-496
• Main Authors: Min, B.J., Korea University, Seoul, Republic of Korea, Kang, S.W., Korea University, Seoul, Republic of Korea, Song, Y.S., Korea University, Seoul, Republic of Korea, Lee, J.H., Korea University, Seoul, Republic of Korea, Lee, S.H., Kwangwoon University, Seoul, Republic of Korea, Park, C.H., Kwangwoon University, Seoul, Republic of Korea, Kim, S.W., Korea University, Seoul, Republic of Korea, Kim, C.W., Korea University, Seoul, Republic of Korea
• Format: Article
• Language: English
• Subjects: acceptance criteria; Anion exchange; anion exchange chromatography; Bioengineering; Biotechnology; Chemistry; Chemistry and Materials Science; CHROMATOGRAPHIE; CHROMATOGRAPHY; CROMATOGRAFIA; Endotoxins; Engineering; Flow rates; Genetic recombination; Growth hormones; Industrial and Production Engineering; Liquid chromatography; Manufacturing; Manufacturing industry; Pituitary gland; Process control; Proteins; Quality control; recombinant human growth hormone (r-hGH); Research Paper; Sodium chloride; Studies; validation; Yeast; 생물공학
• ISSN: 1226-8372; 1976-3816
• DOI: 10.1007/s12257-009-3053-9

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: less than 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: less-than or equal to 0.5 EU/mg, ECP: less-than or equal to 1.4 ppm, IEF: same removal distance, hGH content by Native-PAGE: 100%, purity by HPLC: greater-than or equal to 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.