A MALDI-MS-based quantitative targeted glycomics (MALDI-QTaG) for total N-glycan analysis.
Biotechnol Lett
; 37(10): 2019-25, 2015 Oct.
Article
em En
| MEDLINE
| ID: mdl-26063621
ABSTRACT
OBJECTIVES:
To develop a sensitive and quantitative method for monitoring the abnormal glycosylation of clinical and biopharmaceutical products.RESULTS:
MALDI-MS-based quantitative targeted glycomics (MALDI-QTaG) was proposed for sensitive and quantitative analysis of total N-glycans. The derivatization reactions (i.e., amidation of sialic acid and incorporation of a positive charge moiety into the reducing end) dramatically increased the linearity (R(2) > 0.99) and sensitivity (limit of detection is 0.5 pmol/glycoprotein) relative to underivatized glycans. In addition, the analytical strategy was chromatographic purification-free and non-laborious process accessible to the high-throughput analyses. We used MALDI-QTaG to analyze the N-glycans of α-fetoprotein (AFP) purified from normal cord blood and HCC cell line (Huh7 cells). The total percentages of core-fucosylated AFP N-glycans from Huh7 cells and normal cord blood were 98 and 18%, respectively.CONCLUSIONS:
This MALDI-MS-based glycomics technology has wide applications in many clinical and bioengineering fields requiring sensitive, quantitative and fast N-glycosylation validation.Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Polissacarídeos
/
Glicoproteínas
/
Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
/
Glicômica
Tipo de estudo:
Diagnostic_studies
/
Evaluation_studies
Limite:
Humans
Idioma:
En
Ano de publicação:
2015
Tipo de documento:
Article