Abstract
Liquid chromatography-tandem mass spectrometry-based proteomics for peptide mapping and sequencing was used to characterize the marketed monoclonal antibody trastuzumab and compare it with two biosimilar products, mAb A containing D359E and L361M variations at the Fc site and mAb B without variants. Complete sequence coverage (100%) including disulfide linkages, glycosylations and other commonly occurring modifications (i.e., deamidation, oxidation, dehydration and K-clipping) were identified using maps generated from multi-enzyme digestions. In addition to the targeted comparison for the relative populations of targeted modification forms, a non-targeted approach was used to globally compare ion intensities in tryptic maps. The non-targeted comparison provided an extra-dimensional view to examine any possible differences related to variants or modifications. A peptide containing the two variants in mAb A, D359E and L361M, was revealed using the non-targeted comparison of the tryptic maps. In contrast, no significant differences were observed when trastuzumab was self-compared or compared with mAb B. These results were consistent with the data derived from peptide sequencing via collision induced dissociation/electron transfer dissociation. Thus, combined targeted and non-targeted approaches using powerful mass spectrometry-based proteomic tools hold great promise for the structural characterization of biosimilar products.
| Original language | English |
|---|---|
| Pages (from-to) | 126-135 |
| Number of pages | 10 |
| Journal | Journal of Pharmaceutical and Biomedical Analysis |
| Volume | 80 |
| DOIs | |
| Publication status | Published - 2013 Jun 1 |
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All Science Journal Classification (ASJC) codes
- Analytical Chemistry
- Pharmaceutical Science
- Drug Discovery
- Spectroscopy
- Clinical Biochemistry
Cite this
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A global comparability approach for biosimilar monoclonal antibodies using LC-tandem MS based proteomics. / Chen, Shun Li; Wu, Shiaw Lin; Huang, Li Juan; Huang, Jia Bao; Chen, Shu-Hui.
In: Journal of Pharmaceutical and Biomedical Analysis, Vol. 80, 01.06.2013, p. 126-135.Research output: Contribution to journal › Article
TY - JOUR
T1 - A global comparability approach for biosimilar monoclonal antibodies using LC-tandem MS based proteomics
AU - Chen, Shun Li
AU - Wu, Shiaw Lin
AU - Huang, Li Juan
AU - Huang, Jia Bao
AU - Chen, Shu-Hui
PY - 2013/6/1
Y1 - 2013/6/1
N2 - Liquid chromatography-tandem mass spectrometry-based proteomics for peptide mapping and sequencing was used to characterize the marketed monoclonal antibody trastuzumab and compare it with two biosimilar products, mAb A containing D359E and L361M variations at the Fc site and mAb B without variants. Complete sequence coverage (100%) including disulfide linkages, glycosylations and other commonly occurring modifications (i.e., deamidation, oxidation, dehydration and K-clipping) were identified using maps generated from multi-enzyme digestions. In addition to the targeted comparison for the relative populations of targeted modification forms, a non-targeted approach was used to globally compare ion intensities in tryptic maps. The non-targeted comparison provided an extra-dimensional view to examine any possible differences related to variants or modifications. A peptide containing the two variants in mAb A, D359E and L361M, was revealed using the non-targeted comparison of the tryptic maps. In contrast, no significant differences were observed when trastuzumab was self-compared or compared with mAb B. These results were consistent with the data derived from peptide sequencing via collision induced dissociation/electron transfer dissociation. Thus, combined targeted and non-targeted approaches using powerful mass spectrometry-based proteomic tools hold great promise for the structural characterization of biosimilar products.
AB - Liquid chromatography-tandem mass spectrometry-based proteomics for peptide mapping and sequencing was used to characterize the marketed monoclonal antibody trastuzumab and compare it with two biosimilar products, mAb A containing D359E and L361M variations at the Fc site and mAb B without variants. Complete sequence coverage (100%) including disulfide linkages, glycosylations and other commonly occurring modifications (i.e., deamidation, oxidation, dehydration and K-clipping) were identified using maps generated from multi-enzyme digestions. In addition to the targeted comparison for the relative populations of targeted modification forms, a non-targeted approach was used to globally compare ion intensities in tryptic maps. The non-targeted comparison provided an extra-dimensional view to examine any possible differences related to variants or modifications. A peptide containing the two variants in mAb A, D359E and L361M, was revealed using the non-targeted comparison of the tryptic maps. In contrast, no significant differences were observed when trastuzumab was self-compared or compared with mAb B. These results were consistent with the data derived from peptide sequencing via collision induced dissociation/electron transfer dissociation. Thus, combined targeted and non-targeted approaches using powerful mass spectrometry-based proteomic tools hold great promise for the structural characterization of biosimilar products.
UR - http://www.scopus.com/inward/record.url?scp=84875844715&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84875844715&partnerID=8YFLogxK
U2 - 10.1016/j.jpba.2013.02.040
DO - 10.1016/j.jpba.2013.02.040
M3 - Article
C2 - 23563225
AN - SCOPUS:84875844715
VL - 80
SP - 126
EP - 135
JO - Journal of Pharmaceutical and Biomedical Analysis
JF - Journal of Pharmaceutical and Biomedical Analysis
SN - 0731-7085
ER -