Certification and stability assessment of recombinant human growth hormone as a certified reference material for protein quantification

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 1126-1127; p. 121732
• Main Authors: Tran, Thi Thanh Huong, Kim, Juok, Rosli, Nordiana, Mok, Inkyu, Oh, Kyong Hwa, Lee, Hwashim, Hong, Seon-Pyo, Jin, You-Xun, Wu, Liqing, Wang, Jing, Sakaguchi, Yohei, Kinumi, Tomoya, Takatsu, Akiko, Kim, Sook-Kyung, Jeong, Ji-Seon
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 15.09.2019
• Subjects: Certified reference material; Chromatography, High Pressure Liquid - standards; Human growth hormone; Human Growth Hormone - analysis; Human Growth Hormone - chemistry; Humans; International comparison; Mass Spectrometry - standards; Protein quantification; Protein Stability; Recombinant Proteins - analysis; Recombinant Proteins - chemistry; Reference Standards; Stability assessment; Protein quantification; Certified reference material; Human growth hormone; International comparison; Stability assessment
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2019.121732

A certified reference material (CRM) for the quantification of protein, essential to manage quality control and quality assurance in protein-related works, has been developed. Amino acid analysis with conventional acid hydrolysis and isotope dilution HPLC-MS was used to establish an SI-traceable absolute protein quantification method using recombinant human growth hormone (hGH) as a model protein. The certification method was verified by comparative studies between 1) different methods of protein quantification based on microwave-assisted hydrolysis, and 2) different labs as part of the Asian Collaboration on Reference Material project with Japan, China, and Korea. Certification, evaluation of measurement uncertainty, homogeneity testing, and stability testing were carried out, after which the candidate CRM for hGH quantification was successfully certified with excellent agreement within the certified value in the two comparative studies. Although the quantification value of hGH by amino acid analysis showed good robustness in various conditions, results of intact protein analysis showed degradation profiles in temperatures higher than 4 °C. Consequently, storage and dissemination conditions should be set in accordance with stability tests. Based on the results, this method is believed to be suitable for accurate quantification of hGH. Additionally, it can also be used as a guide to preparation of CRM, and instructions for quality management of protein work for other similar proteins. •Full certification process of reference material for protein quantification using human growth hormone (hGH) as a model.•An establishment of reference measurement procedure for absolute quantification of hGH using isotope-dilution mass spectrometry (ID-MS)•Assessment of the quantification accuracy of the proposed method using several methods of protein quantification, and inter-laboratory comparison as well.•The most straightforward approach for SI-traceable protein quantification•Strict investigation of stability concerns on protein as reference material