In:
Rapid Communications in Mass Spectrometry, Wiley, Vol. 28, No. 8 ( 2014-04-30), p. 855-860
Abstract:
Host cell proteins (HCPs), which are process‐related impurities typically present at low levels in recombinant biopharmaceutical products, are often measured using an immunological technique, such as an enzyme‐linked immunosorbent assay (ELISA). In contrast to ELISA which only provides the total amount of HCP, liquid chromatography/mass spectrometry (LC/MS) can provide both qualitative and quantitative information about the major HCP species. In this study, an HCP‐enrichment step was optimized and combined with LC/MS to identify and determine the relative abundance of HCPs present in a monoclonal antibody (mAb) drug product. METHODS An NS0 (mouse myeloma) cell‐derived mAb drug product, whose total HCP level was less than 100 ng/mg of protein, was subjected to analysis by LC/MS. One‐dimensional and two‐dimensional chromatography options, together with the off‐line HCP enrichment strategy based on Protein A chromatography, were evaluated for optimal HCP detection. RESULTS With this approach, nineteen HCPs were detected from a therapeutic mAb, an improvement over the detection of only one HCP without depletion. CONCLUSIONS Compared with other published HCP studies with LC/MS, the HCP‐enrichment step in our method enables a more practical and relevant application to approved protein therapeutics, which are mostly mammalian cell‐derived products with HCPs present at very low levels. Copyright © 2014 John Wiley & Sons, Ltd.
Type of Medium:
Online Resource
ISSN:
0951-4198
,
1097-0231
Language:
English
Publisher:
Wiley
Publication Date:
2014
detail.hit.zdb_id:
2002158-6
detail.hit.zdb_id:
58731-X
SSG:
11
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