Application of middle-down approach in quantitation and catabolite identification of protein by LC-high-resolution mass spectrometry.
Bioanalysis
; 13(6): 465-479, 2021 Mar.
Article
de En
| MEDLINE
| ID: mdl-33719526
ABSTRACT
Aim:
To further enhance the detection sensitivity and increase resolving power of top-down intact protein bioanalysis, middle-down approach was explored. Materials &methods:
An monoclonal antibody (mAb) was used as a model protein to evaluate quantitative bioanalytical assay performance and a disulfide linked dimer protein was investigated for its pharmacokinetics properties and catabolism in vivo by middle-down approach. Results &Conclusion:
For quantitation of the mAb, different subunits generated by middle-down approach provided different level of signal improvement in biological samples with Lc and half Fc giving five-times better sensitivity than intact mAb. For the dimer protein, middle-down analysis by reduction enabled effective differentiation of the unchanged protein and its oxidized form, and clearly elucidated their respective proteolytic catabolites.Mots clés
Texte intégral:
1
Collection:
01-internacional
Base de données:
MEDLINE
Sujet principal:
Anticorps monoclonaux
Type d'étude:
Diagnostic_studies
Limites:
Humans
Langue:
En
Journal:
Bioanalysis
Année:
2021
Type de document:
Article
Pays d'affiliation:
États-Unis d'Amérique