Development of a liquid chromatography-based versatile bioanalysis for bevacizumab based on pretreatment combining aptamer affinity purification and centrifugal ultrafiltration concentration.
Kenichiro TodorokiDaichi HamadaTomohiro YamadaTaro SaitoYutaka ShimizuEiji SugiyamaHajime MizunoHideki HayashiKaori TsukakoshiKazunori IkebukuroPublished in: Analytical sciences : the international journal of the Japan Society for Analytical Chemistry (2023)
We report on the development of a versatile and accurate bioanalytical method for bevacizumab using a pretreatment method combining affinity purification with anti-idiotypic DNA aptamers and centrifugal ultrafiltration concentration, followed by liquid chromatography (LC)-fluorescence analysis. An affinity purification method using Sepharose beads as an affinity support removed immunoglobulin G and a large amount of coexisting substances in the serum sample. Purified bevacizumab was separated as a single peak by conventional LC and detected fluorometrically, showing good linearity (R 2 = 0.999) in the range of 5-200 μg/mL, sufficient to analyze bevacizumab concentrations in the blood of bevacizumab-treated patients. By combining this purification method with a concentration method using a centrifugal filtration device that inhibits non-specific adsorption of bevacizumab, the quantitative range was reduced by a factor of 10 while showing good linearity (R 2 = 0.999) in the 0.5-20 μg/mL range. The developed analytical method is expected to be used not only for general bioanalysis of therapeutic mAbs in clinical settings, but also for next-generation antibody drugs that show drug efficacy at low concentrations and for analysis of trace samples.
Keyphrases
- liquid chromatography
- metastatic colorectal cancer
- mass spectrometry
- end stage renal disease
- emergency department
- high resolution mass spectrometry
- chronic kidney disease
- newly diagnosed
- high resolution
- ejection fraction
- capillary electrophoresis
- prognostic factors
- risk assessment
- peritoneal dialysis
- circulating tumor cells
- recombinant human
- drinking water
- patient reported outcomes
- sensitive detection
- adverse drug
- circulating tumor
- quantum dots