High-sensitivity quantitation of a Nanobody® in plasma by single-cartridge multidimensional SPE and ultra-performance LC-MS/MS.
Bioanalysis
; 7(1): 53-64, 2015.
Article
in En
| MEDLINE
| ID: mdl-25558935
ABSTRACT
BACKGROUND:
A major challenge in protein quantitation based on enzymatic digestion of complex biological samples and subsequent LC-MS/MS analysis of a signature peptide is dealing with the high complexity of the matrix after digestion, which can reduce sensitivity considerably.RESULTS:
Using single cartridge multidimensional SPE, sufficient selectivity was introduced to allow quantitation in 50 µl of plasma down to 10.0 ng/ml (~0.3 nM). An inhouse prepared (18)O-labeled signature peptide was used as the internal standard. The procedure was validated for human and rabbit plasma.CONCLUSION:
The developed SPE procedure allowed the sensitive and selective LC-MS/MS quantitation of the Nanobody(®) without the use of antibodies. When appropriate precautions are taken, the (18)O-labeled peptide is a practical and economical alternative to custom synthesis.Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Mass Spectrometry
/
Chromatography, Liquid
/
Single-Domain Antibodies
Type of study:
Diagnostic_studies
Limits:
Humans
Language:
En
Journal:
Bioanalysis
Year:
2015
Document type:
Article
Affiliation country:
Netherlands