Anti-VEGF Drug Interference with Vegf Quantitation in the R&amp;D Systems Human Quantikine VEGF ELISA Kit

Sumner, Giane; Georgaros, Camille; Rafique, Ashique; DiCioccio, Thomas; Martin, Joel; Papadopoulos, Nicholas; Daly, Thomas J.; Torri, Albert

doi:10.4155/bio-2018-0096

Cited by 22 publications

(16 citation statements)

References 16 publications

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“…The second set of experiments examined the ability of bevacizumab to inhibit VEGFA secretion in vitro at normoxic and hypoxic conditions. Although bevacizumab could compete with the detection antibody, ELISA assay is able to measure the total VEGFA present in the extracellular media [29]. For this purpose, we used a standard dose (SD; 8.3 μg/ml) which mimics the clinical administration and a specific dose (Spe) calculated for each cell line (see Additional file 1: methods section).…”

Section: Specific But Not Standard Bevacizumab Doses Impair Vegfa Expression and Secretionmentioning

confidence: 99%

Bevacizumab dose adjustment to improve clinical outcomes of glioblastoma

García-Romero

Palacín-Aliana

Madurga

et al. 2020

BMC Med

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Background: Glioblastoma (GBM) is one of the most aggressive and vascularized brain tumors in adults, with a median survival of 20.9 months. In newly diagnosed and recurrent GBM, bevacizumab demonstrated an increase in progression-free survival, but not in overall survival. Methods: We conducted an in silico analysis of VEGF expression, in a cohort of 1082 glioma patients. Then, to determine whether appropriate bevacizumab dose adjustment could increase the anti-angiogenic response, we used in vitro and in vivo GBM models. Additionally, we analyzed VEGFA expression in tissue, serum, and plasma in a cohort of GBM patients before and during bevacizumab treatment.Results: We identified that 20% of primary GBM did not express VEGFA suggesting that these patients would probably not respond to bevacizumab therapy as we proved in vitro and in vivo. We found that a specific dose of bevacizumab calculated based on VEGFA expression levels increases the response to treatment in cell culture and serum samples from mice bearing GBM tumors. Additionally, in a cohort of GBM patients, we observed a correlation of VEGFA levels in serum, but not in plasma, with bevacizumab treatment performance. Conclusions: Our data suggest that bevacizumab dose adjustment could improve clinical outcomes in Glioblastoma treatment.

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Section: Specific But Not Standard Bevacizumab Doses Impair Vegfa Expression and Secretionmentioning

confidence: 99%

Bevacizumab dose adjustment to improve clinical outcomes of glioblastoma

García-Romero

Palacín-Aliana

Madurga

et al. 2020

BMC Med

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“…The current minimum detectable aflibercept concentration is 0.2 µg/mL, which is almost equal to the calculated aqueous aflibercept concentration 2 months after injection. 19 , 28 Therefore, the duration of aflibercept efficacy may not be compromised, even if aflibercept leakage is substantial. Hence, the current method is a valuable tool to assess pharmacokinetics in the eye.…”

Section: Discussionmentioning

confidence: 99%

Microvolume Analysis of Aflibercept in Aqueous Humor Using Mass Spectrometry

Kimura

Takahashi

Sakamoto

et al. 2022

Trans. Vis. Sci. Tech.

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Purpose To develop a microvolume analytical method for measurement of the aflibercept concentration in human intraocular fluid and plasma. Methods We analyzed trace amounts of aflibercept in human aqueous humor using Fab-selective proteolysis and nano-surface and molecular-orientation limited (nSMOL) proteolysis, coupled with liquid chromatography–tandem mass spectrometry (LC-MS/MS). Patients with age-related macular degeneration or diabetic macular edema were recruited. Just after an injection of 50 µL of aflibercept, regurgitate from needle holes was collected with a micropipette pressed to the side of the injection hole within 10 seconds. The median amount of regurgitate was 4 µL (range, 1–18 µL). Results In human plasma, the aflibercept concentration ranged between 0.195 and 50 µg/mL when using the quantitative signature peptide IIWDSR (aa. 56–61) present on the vascular endothelial growth factor receptor 1 domain of aflibercept. The method was validated by evaluating its linearity, carryover, selectivity, accuracy and precision, dilution effect, and sample/processing stability. As only a minimal amount of regurgitate through needle holes can be sampled, we performed and verified the aflibercept assay using patient samples after 1:10 dilution with control human plasma, a recognized diluent. The median concentration of aflibercept in the regurgitate was 240 µg/mL (range, 13–4300 µg/mL). Conclusions Our findings indicate that the aflibercept assay using human intraocular fluid can be reliably performed using nSMOL coupled with LC-MS/MS. Translational Relevance This technique for quantifying aflibercept in the regurgitate suggests that the amount of drug lost post-injection can be ignored, even in patients with a relatively large leak after vitreous injection. This new methodology suggests possible therapeutic responses and may be employed as a general analytical method for trapping many biologics, such as vascular endothelial growth factor, in various types of clinical samples, unaffected by proteinaceous or small organic pharmaceuticals.

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“…The aforesaid concentration is consistent to the peak of serum concentration of Aflibercept (Cmax) in vivo and after the first intravitreal injection would mimics its systemic effect reasonably. Cmax is obtained by investigating recent anti–VEGF‐associated studies that evaluated the expression level of specific genes on vascular endothelial cells 47‐52 . RNA extraction was performed from HUVEC cells that had been treated 6 hours or 24 hours by Aflibercept (Bayer, Germany) or phosphate‐buffered saline (PBS, as a control).…”

Section: Methodsmentioning

confidence: 99%

Network analysis and the impact of Aflibercept on specific mediators of angiogenesis in HUVEC cells

Latifi-Navid

Soheili

Samiei

et al. 2021

J Cellular Molecular Medi

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Angiogenesis, inflammation and endothelial cells’ migration and proliferation exert fundamental roles in different diseases. However, more studies are needed to identify key proteins and pathways involved in these processes. Aflibercept has received the approval of the US Food and Drug Administration (FDA) for the treatment of wet AMD and colorectal cancer. Moreover, the effect of Aflibercept on VEGFR2 downstream signalling pathways has not been investigated yet. Here, we integrated text mining data, protein‐protein interaction networks and multi‐experiment microarray data to specify candidate genes that are involved in VEGFA/VEGFR2 signalling pathways. Network analysis of candidate genes determined the importance of the nominated genes via different centrality parameters. Thereupon, several genes—with the highest centrality indexes—were recruited to investigate the impact of Aflibercept on their expression pattern in HUVEC cells. Real‐time PCR was performed, and relative expression of the specific genes revealed that Aflibercept modulated angiogenic process by VEGF/PI3KA/AKT/mTOR axis, invasion by MMP14/MMP9 axis and inflammation‐related angiogenesis by IL‐6‐STAT3 axis. Data showed Aflibercept simultaneously affected these processes and determined the nominated axes that had been affected by the drug. Furthermore, integrating the results of Aflibercept on expression of candidate genes with the current network analysis suggested that resistance against the Aflibercept effect is a plausible process in HUVEC cells.

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Anti-VEGF Drug Interference with Vegf Quantitation in the R&D Systems Human Quantikine VEGF ELISA Kit

Cited by 22 publications

References 16 publications

Bevacizumab dose adjustment to improve clinical outcomes of glioblastoma

Bevacizumab dose adjustment to improve clinical outcomes of glioblastoma

Microvolume Analysis of Aflibercept in Aqueous Humor Using Mass Spectrometry

Network analysis and the impact of Aflibercept on specific mediators of angiogenesis in HUVEC cells

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