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AFLIBERCEPT ELISA

AFLIBERCEPT ELISA

SKU: AA115
Enzyme immunoassay for the quantitative determination of free Aflibercept (Eylea®, Zaltrap®) in serum and plasma.
The solid phase (MTP) is coated by the target molecule, in this case VEGF.
 
Required Volume (µL)10
Incubation Time (min)105
SampleSerum or Plasma
Plate Size96 Tests
Standard Range (ng/mL, 10x)0-2000
Detection Limit (ng/mL)5
Spike Recovery (%)>95
Shelf Life (years)2
 
Intended Use: This kit has been developed for the measurement of drug levels in research, diagnostic and biosimilar uses. It is suitable for Therapeutical Drug Monitoring (TDM) purposes.
 
Eylea® is a trademark of Bayer.
Zaltrap® is a trademark of Sanofi Genzyme.

  • ESSAY CHARACTERISTICS

    SPECIFICITY

    There is no cross reaction with any other proteins present in native human serum. A screening test was performed with 48 different native human sera. All produced OD450/620 nm values less than the mean OD of standard D. No cross reaction was observed with the other therapeutic antibodies tested (Tocilizumab, Cetuximab, Etanercept, Infliximab, Rituximab and Trastuzumab) at concentrations up to 2 mg/mL. However, Bevacizumab resulted in full cross-reaction as expected. Ranibizumab, another anti-VEGF therapeutic antibody consisted of Fab fragment only, produced a mean OD less than the mean OD of standard D.

     

    SENSITIVITY

    The lowest detectable level that can be clearly distinguished from the zero standard is 5 ng/mL (zero standard +2SD read from the curve) under the above-described conditions. Analytical sensitivity is 5 ng/mL, and corresponding to the detection limit (limit of quantification) of 0.5 µg/mL for undiluted clinical samples because the serum or plasma samples are instructed to be diluted at 1:100 before starting the assay

     

    PRECISION

    Intra-assay CV: <10%.

    Inter-assay CV: <10%.

     

    RECOVERY

    Recovery rate was found to be >95% with native serum and plasma samples when spiked with exogenous Aflibercept.

     

    AUTOMATION

    The ImmunoGuide Aflibercept ELISA is suitable also for being used by an automated ELISA processor.

  • REFERENCES

    1. Park SJ, Choi Y, Na YM, Hong HK, Park JY, Park KH, Chung JY, Woo SJ. Intraocular Pharmacokinetics of Intravitreal Aflibercept (Eylea) in a Rabbit Model. Invest Ophthalmol Vis Sci. 2016;57(6):2612-7.

    2. Stewart MW1, Rosenfeld PJ, Penha FM, Wang F, Yehoshua Z, Bueno-Lopez E, Lopez PF. Pharmacokinetic rationale for dosing every 2 weeks versus 4 weeks with intravitreal ranibizumab, bevacizumab, and aflibercept (vascular endothelial growth factor Trap-eye). Retina. 2012;32(3):434-57.

    3. Arnott C, Punnia-Moorthy G, Tan J, Sadeghipour S, Bursill C, Patel S. The Vascular Endothelial Growth Factor Inhibitors Ranibizumab and Aflibercept Markedly Increase Expression of Atherosclerosis-Associated Inflammatory Mediators on Vascular Endothelial Cells. PLoS One. 2016;11(3):e0150688. doi: 10.1371.

    4. Takahashi H, Nomura Y, Nishida J, Fujino Y, Yanagi Y, Kawashima H. Vascular Endothelial Growth Factor (VEGF) Concentration Is Underestimated by Enzyme-Linked Immunosorbent Assay in the Presence of Anti-VEGF Drugs. Invest Ophthalmol Vis Sci. 2016;57(2):462-6.

    5. Puddu A, Sanguineti R, Traverso CE, Viviani GL, Nicolò M. Response to anti-VEGF-A treatment of endothelial cells in vitro. Exp Eye Res. 2016;146:128-36.

    6. Niwa Y, Kakinoki M, Sawada T, Wang X, Ohji M. Ranibizumab and Aflibercept: Intraocular Pharmacokinetics and Their Effects on Aqueous VEGF Level in Vitrectomized and Nonvitrectomized Macaque Eyes. Invest Ophthalmol Vis Sci. 2015;56(11):6501-5.

    7. Celik N, Scheuerle A, Auffarth GU, Kopitz J, Dithmar S. Intraocular Pharmacokinetics of Aflibercept and Vascular Endothelial Growth Factor-A. Invest Ophthalmol Vis Sci. 2015;56(9):5574-8.

    8. Giurdanella G, Anfuso CD, Olivieri M, Lupo G, Caporarello N, Eandi CM, Drago F, Bucolo C, Salomone S. Aflibercept, bevacizumab and ranibizumab prevent glucose-induced damage in human retinal pericytes in vitro, through a PLA2/COX-2/VEGF-A pathway. Biochem Pharmacol. 2015;96(3):278-87.

    9. Zehetner C, Bechrakis NE, Stattin M, Kirchmair R, Ulmer H, Kralinger MT, Kieselbach GF. Systemic counterregulatory response of placental growth factor levels to intravitreal aflibercept therapy. Invest Ophthalmol Vis Sci. 2015;56(5):3279-86.

    10. Klettner A, Grotelüschen S, Treumer F, Roider J, Hillenkamp J. Compatibility of recombinant tissue plasminogen activator (rtPA) and aflibercept or ranibizumab coapplied for neovascular age-related macular degeneration with submacular haemorrhage. Br J Ophthalmol. 2015;99(6):864-9.

    11. Mansour AM, Al-Ghadban SI, Yunis MH, El-Sabban ME. Ziv-aflibercept in macular disease. Br J Ophthalmol. 2015;99(8):1055-9.

    12. Park SJ, Oh J, Kim YK, Park JH, Park JY, Hong HK, Park KH, Lee JE, Kim HM, Chung JY, Woo SJ. Intraocular pharmacokinetics of intravitreal vascular endothelial growth factor-Trap in a rabbit model. Eye (Lond). 2015;29(4):561-8.

    13. Zehetner C, Kralinger MT, Modi YS, Waltl I, Ulmer H, Kirchmair R, Bechrakis NE, Kieselbach GF. Systemic levels of vascular endothelial growth factor before and after intravitreal injection of aflibercept or ranibizumab in patients with age-related macular degeneration: a randomised, prospective trial. Acta Ophthalmol. 2015;93(2):e154-9. doi: 10.1111/aos.12604.

    14. Klettner A, Recber M, Roider J. Comparison of the efficacy of aflibercept, ranibizumab, and bevacizumab in an RPE/choroid organ culture. Graefes Arch Clin Exp Ophthalmol. 2014;252(10):1593-8.

    15. Yoshida I, Shiba T, Taniguchi H, Takahashi M, Murano T, Hiruta N, Hori Y, Bujo H, Maeno T. Evaluation of plasma vascular endothelial growth factor levels after intravitreal injection of ranibizumab and aflibercept for exudative age-related macular degeneration. Graefes Arch Clin Exp Ophthalmol. 2014;252(9):1483-9.

    16. Avery RL, Castellarin AA, Steinle NC, Dhoot DS, Pieramici DJ, See R, Couvillion S, Nasir MA, Rabena MD, Le K, Maia M, Visich JE. Systemic pharmacokinetics following intravitreal injections of ranibizumab, bevacizumab or aflibercept in patients with neovascular AMD. Br J Ophthalmol. 2014;98(12):1636-41.

    17. Wang X, Sawada T, Sawada O, Saishin Y, Liu P, Ohji M. Serum and plasma vascular endothelial growth factor concentrations before and after intravitreal injection of aflibercept or ranibizumab for age-related macular degeneration. Am J Ophthalmol. 2014;158(4):738-744.

    18. Shonka N, Piao Y, Gilbert M, Yung A, Chang S, DeAngelis LM, Lassman AB, Liu J, Cloughesy T, Robins HI, Lloyd R, Chen A, Prados M, Wen PY, Heymach J, de Groot J. Cytokines associated with toxicity in the treatment of recurrent glioblastoma with aflibercept. Target Oncol. 2013;8(2):117-25.

    19. de Groot JF, Piao Y, Tran H, Gilbert M, Wu HK, Liu J, Bekele BN, Cloughesy T, Mehta M, Robins HI, Lassman A, DeAngelis L, Camphausen K, Chen A, Yung WK, Prados M, Wen PY, Heymach JV. Myeloid biomarkers associated with glioblastoma response to anti-VEGF therapy with aflibercept. Clin Cancer Res. 2011;17(14):4872-81.

    20. Lassoued W, Murphy D, Tsai J, Oueslati R, Thurston G, Lee WM. Effect of VEGF and VEGF Trap on vascular endothelial cell signaling in tumors. Cancer Biol Ther. 2010;10(12):1326-33.

  • INSTRUCTIONS FOR USE

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  • SAFETY DATA SHEET

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  • BATCH/LOT INFORMATION

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