A novel LC-MS/MS approach to the pharmacokinetic study of free and bound aflibercept simultaneously.
Anal Bioanal Chem
; 412(4): 1003-1010, 2020 Feb.
Article
in En
| MEDLINE
| ID: mdl-31897562
ABSTRACT
To comprehensively evaluate the pharmacokinetic (PK) characteristics of aflibercept, we established a liquid chromatography with tandem mass spectrometry (LC-MS/MS) method to determine the concentration of vascular endothelial growth factor (VEGF)-A-bound aflibercept and free aflibercept. A specific sample preparation method of nano-surface and molecular-orientation limited (nSMOL) proteolysis was performed to extract both free and bound aflibercept from plasma. The tryptic peptides unique to aflibercept and VEGF-A were selected to quantify the amounts of total aflibercept and aflibercept-VEGF complex, respectively. The method was validated by evaluating its selectivity, linearity, precision, accuracy, extraction recovery, matrix effect, and stability. It was then successfully used to quantify total and bound aflibercept concentrations in cynomolgus monkey plasma, while indirectly obtaining the concentration of free aflibercept by subtraction. The PK results of this LC-MS/MS method are comparable to the traditional enzyme-linked immunosorbent assay (ELISA) results. It is thus a reliable and complementary method for the PK evaluation of aflibercept. Graphical abstract.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Recombinant Fusion Proteins
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Chromatography, High Pressure Liquid
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Receptors, Vascular Endothelial Growth Factor
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Vascular Endothelial Growth Factor A
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Tandem Mass Spectrometry
Type of study:
Evaluation_studies
Limits:
Animals
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Humans
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Male
Language:
En
Journal:
Anal Bioanal Chem
Year:
2020
Document type:
Article