Anti-Idiotype DNA Aptamer Affinity Purification⁻High-Temperature Reversed-Phase Liquid Chromatography: A Simple, Accurate, and Selective Bioanalysis of Bevacizumab.
Tomohiro YamadaTaro SaitoYutaka ShimizuKaori TsukakoshiHideki HayashiHajime MizunoDaiki TsujiKeisuke YamamotoKunihiko ItohToshimasa Toyo'okaKazunori IkebukuroKenichiro TodorokiPublished in: Molecules (Basel, Switzerland) (2019)
This study presents a simple, accurate, and selective bioanalytical method of bevacizumab detection from plasma samples based on aptamer affinity purification⁻high-temperature reversed-phased liquid chromatography (HT-RPLC) with fluorescence detection. Bevacizumab in plasma samples was purified using magnetic beads immobilized with an anti-idiotype DNA aptamer for bevacizumab. The purified bevacizumab was separated with HT-RPLC and detected with its native fluorescence. Using aptamer affinity beads, bevacizumab was selectively purified and detected as a single peak in the chromatogram. HT-RPLC achieved good separation for bevacizumab with a sharp peak within 10 min. The calibration curves of the two monoclonal antibodies ranged from 1 to 50 μg/mL and showed good correlation coefficients (r² > 0.999). The limit of detection (LOD) and lower limit of quantification (LLOQ) values for bevacizumab were 0.15 and 0.51 μg/mL, respectively. The proposed method was successfully applied to the bioanalysis of the plasma samples obtained from the patients with lung cancer and may be extended to plan optimal therapeutic programs and for the evaluation of biological equivalencies in the development of biosimilars.
Keyphrases
- metastatic colorectal cancer
- liquid chromatography
- high temperature
- label free
- gold nanoparticles
- mass spectrometry
- sensitive detection
- magnetic nanoparticles
- public health
- loop mediated isothermal amplification
- high resolution mass spectrometry
- real time pcr
- tandem mass spectrometry
- cell free
- simultaneous determination
- high resolution
- nucleic acid
- solid phase extraction
- molecularly imprinted