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Old 08-04-2019, 06:01 AM
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rss Human Growth Hormone - Certification and stability assessment of recombinant human gr

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

J Chromatogr B Analyt Technol Biomed Life Sci. 2019 Jul 25;1126-1127:121732

Authors: Tran TTH, Kim J, Rosli N, Mok I, Oh KH, Lee H, Hong SP, Jin YX, Wu L, Wang J, Sakaguchi Y, Kinumi T, Takatsu A, Kim SK, Jeong JS

Abstract
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.


PMID: 31376580 [PubMed - as supplied by publisher]



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