Therapeutic Drug Monitoring ELISA Assay

Trastuzumab mAb-based ELISA Assay

$1,485.00

The Trastuzumab mAb-based ELISA Assay is an enzyme immunoassay for the specific and precise determination of free Trastuzumab in serum and plasma. The Trastuzumab (mAb-based) ELISA Assay Kit is for research use only and not to be used in diagnostic procedures.

SKU: IG-AB105 Categories: , , , Tags: , ,

Trastuzumab mAb-based ELISA Assay

The Trastuzumab mAb-based ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 2 ng/mL
Dynamic Range: 6 – 200 ng/mL
Incubation Time: 2 hours
Sample Type: Serum, Plasma
Sample Size: 10 µL
Alternative Names: Herceptin, Herclon

Assay Background

The drug Trastuzumab (trade names Herceptin® or Herclon®) is a recombinant DNA-derived humanized monoclonal antibody that selectively targets the extracellular domain of the human epidermal growth factor receptor 2 protein (HER2). The antibody is an IgG1 kappa that contains human framework regions with the complementarity-determining regions of a murine anti-p185 HER2 antibody that binds to HER2. Trastuzumab blood concentrations throughout the dosing interval expected to be remaining above those considered necessary for anticancer activity. Furthermore, in a separate analysis, patients with the lowest Trastuzumab serum trough concentrations had the highest rate of disease progression and shortest overall survival. This Eagle Biosciences ImmunoGuide Trastuzumab ELISA (mAb-based) is developed for the specific measurement of Trastuzumab in serum, plasma,and other biological fluids by the advantage of using a site-directed VME-5A6b mouse monoclonal antibody (mAb) specific for Trastuzumab only.

STORAGE AND STABILITY
The kit is shipped at ambient temperature and should be stored at 2-8°C. Keep away from heat or direct sun light. The microtiter strips are stable up to the expiry date of the kit in the broken, but tightly closed bag when stored at 2–8°C.

Related Products

Omalizumab mAb Based ELISA Assay
Cetuximab (Erbitux) (mAb Based) ELISA

Additional Information

Assay Priniciple

This Eagle Biosciences ELISA Assay Kit is based on Trastuzumab-specific mouse monoclonal antibody (catcher Ab, ImmunoGuide clone VME-5A6b). Diluted standards and samples are incubated in the microtiter plate coated with IG-VME-5A6b mAb. After incubation, the wells are washed. A horseradish peroxidase (HRP)-conjugated anti-human IgG monoclonal antibody is added and binds to the Fc part of Trastuzumab. Following incubation, wells are washed and the bound enzymatic activity is detected by addition of chromogen-substrate. The color developed is proportional to the amount of Trastuzumab in the sample or standard. Results of samples can be determined by using the standard curve. Binding of Trastuzumab to the solid phase, pre-coated with VME-5A6b, is inhibited by recombinant human HER2 protein in a concentration dependent manner. Therefore, the ImmunoGuide Trastuzumab ELISA (mAb-Based) measures the free form of Trastuzumab.

Assay Procedure

  1. Pipette 100 µL of Assay Buffer into each of the wells to be used.
  2. Pipette 75 µL of each 1:10 Diluted Standard, and 1:1000 Diluted Samples into the respective wells of the microtiter plate.
  3. Cover the plate with adhesive seal. Shake plate carefully. Incubate 60 min at room temperature (RT, 20-25°C).
  4. Remove adhesive seal. Aspirate or decant the incubation solution. Wash the plate 3 X 300 μL of Diluted Wash Buffer per well. Remove excess solution by tapping the inverted plate on a paper towel.
  5. Pipette 100 μL of Enzyme Conjugate (HRP-anti human IgG mAb) into each well.
  6. Cover plate with adhesive seal. Shake plate carefully. Incubate 30 min at RT.
  7. Remove adhesive seal. Aspirate or decant the incubation solution. Wash the plate 3 X 300 μL of Diluted Wash Buffer per well. Remove excess solution by tapping the inverted plate on a paper towel.
  8. Pipette 100 µL of Ready-to-Use TMB Substrate Solution into each well.
  9. Incubate 10 min at RT. Avoid exposure to direct sunlight.
  10. Stop the substrate reaction by adding 100 µL of Stop Solution into each well.
  11. Briefly mix contents by gently shaking the plate. Color changes from blue to yellow.
  12. Measure optical density (OD) with a photometer at 450 nm (Reference at OD620 nm is optional) within 15 min after pipetting the Stop Solution.

Typical Standard Curve

Specificity

There is no cross reaction with any other proteins present in native human serum. A screening test was performed with 36 different native human sera. All produced OD450/620 nm values less than the mean OD of standard D (6 ng/mL). No cross reaction was observed with sera spiked with the other therapeutic antibodies including Infliximab, Adalimumab, Etanercept, Rituximab, Tocilizumab, Omalizumab and Bevacizumab at concentrations up to 2mg/mL. All produced mean OD450/620 nm values less than the mean OD of standard D (6 ng/mL). In addition, binding of Trastuzumab is inhibited by recombinant human HER2 in a concentration dependent manner. Therefore, the ImmunoGuide Trastuzumab ELISA (mAb-based) measures the biologically active free form of Trastuzumab, i.e. not pre-occupied by HER2.

Documents

Product Manual

 

Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.

Publications

References

  • Kang YK, Rha SY, Tassone P, Barriuso J, Yu R, Szado T, Garg A, Bang YJ. A phase IIa dose-finding and safety study of first-line pertuzumab in combination with trastuzumab, capecitabine and cisplatin in patients with HER2-positive advanced gastric cancer. Br J Cancer. 2014;111(4):660-6.
  • Sanford M. Subcutaneous trastuzumab: a review of its use in HER2-positive breast cancer. Target Oncol. 2014 Mar;9(1):85-94.
  • Ismael G, Hegg R, Muehlbauer S, Heinzmann D, Lum B, Kim SB, Pienkowski T, Lichinitser M, Semiglazov V, Melichar B, Jackisch C. Subcutaneous versus intravenous administration of (neo)adjuvant trastuzumab in patients with HER2-positive, clinical stage I-III breast cancer (HannaH study): a phase 3, open-label, multicentre, randomised trial. Lancet Oncol. 2012;13(9):869-78.
  • Leyland-Jones B, Colomer R, Trudeau ME, Wardley A, Latreille J, Cameron D, Cubedo R, Al-Sakaff N, Feyereislova A, Catalani O, Fukushima Y, Brewster M, Cortés J. Intensive loading dose of trastuzumab achieves higher-than-steady-state serum concentrations and is well tolerated. J Clin Oncol. 2010;28(6):960-6.
  • Baselga J, Carbonell X, Castañeda-Soto NJ, Clemens M, Green M, Harvey V, Morales S, Barton C, Ghahramani P. Phase II study of efficacy, safety, and pharmacokinetics of trastuzumab monotherapy administered on a 3-weekly schedule. J Clin Oncol. 2005;23(10):2162-71.
  • Hourcade-Potelleret F, Lemenuel-Diot A, McIntyre C, Brewster M, Lum B, Bittner B. Use of a population pharmacokinetic approach for the clinical development of a fixed-dose subcutaneous formulation of trastuzumab. CPT Pharmacometrics Syst Pharmacol. 2014 Jan 2;3:e87. doi: 10.1038/psp.2013.63.
  • Tokuda Y, Watanabe T, Omuro Y, Ando M, Katsumata N, Okumura A, Ohta M, Fujii H, Sasaki Y, Niwa T, Tajima T. Dose escalation and pharmacokinetic study of a humanized anti-HER2 monoclonal antibody in patients with HER2/neu-overexpressing metastatic breast cancer. Br J Cancer. 1999;81(8):1419-25.
  • Goldenberg MM. Trastuzumab, a recombinant DNA-derived humanized monoclonal antibody, a novel agent for the treatment of metastatic breast cancer. Clin Ther. 1999;21(2):309-18.
  • Arpino G, Michelotti A, Truini M, Montemurro F, Russo S, Palumbo R, Zamagni C, Latorre A, Bruzzese D, Riccardi F, De Laurentiis M, Beano A, Biganzoli L, Zaniboni A, Laudadio L, Malagoli M, Bilancia D, Schettini F, Giuliano M, Cazzaniga ME, De Placido S. Demographic, tumor and clinical features of clinical trials versus clinical practice patients with HER2-positive early breast cancer: results of a prospective study. J Cancer Res Clin Oncol. 2015 Aug 25. [Epub ahead of print]
  • Laboissiere RS, Buzelin MA, Balabram D, De Brot M, Nunes CB, Rocha RM, Cabral MM, Gobbi H. Association between HER2 status in gastric cancer and clinicopathological features: a retrospective study using whole-tissue sections. BMC Gastroenterol. 2015;15(1):157.
  • Kim YS, Sym SJ, Baek MY, Park I, Hong J, Ahn HK, Park J, Cho EK, Lee WK, Chung M, Kim HS, Lee JH, Shin DB. Low-dose capecitabine plus trastuzumab as first-line treatment in patients 75 years of age or older with HER2-positive advanced gastric cancer: a pilot study. Cancer Chemother Pharmacol. 2015 Oct 19. [Epub ahead of print]
  • Martin-Castillo B, Lopez-Bonet E, Cuyàs E, Viñas G, Pernas S, Dorca J, Menendez JA. Cancer stem cell-driven efficacy of trastuzumab (Herceptin): towards a reclassification of clinically HER2-positive breast carcinomas. Oncotarget. 2015;6(32):32317-38.
  • Xu W, Bi Y, Zhang J, Kong J, Jiang H, Tian M, Li K, Wang B, Chen C, Song F, Pan X, Shi B, Kong X, Gu J, Cai X, Li Z. Synergistic antitumor efficacy against the EGFRvIII+HER2+ breast cancers by combining trastuzumab with anti-EGFRvIII antibody CH12. Oncotarget. 2015 Oct 14. doi: 10.18632/oncotarget.6111. [Epub ahead of print]
  • Yeo B, Kotsori K, Mohammed K, Walsh G, Smith IE. Long-term outcome of HER2 positive metastatic breast cancer patients treated with first-line trastuzumab. Breast. 2015 Oct 8. pii: S0960-9776(15)00209-X. doi: 10.1016/j.breast.2015.09.008.
  • D’Avino C, Paciello R, Riccio G, Coppola C, Laccetti P, Maurea N, Raines RT, De Lorenzo C. Effects of a second-generation human anti-ErbB2 ImmunoRNase on trastuzumab-resistant tumors and cardiac cells. Protein Eng Des Sel. 2014;27(3):83-8.
  • Clay TM, Osada T, Hartman ZC, Hobeika A, Devi G, Morse MA, Lyerly HK. Polyclonal immune responses to antigens associated with cancer signaling pathways and new strategies to enhance cancer vaccines. Immunol Res. 2011;49(1-3):235-47
  • Wong JY, Raubitschek A, Yamauchi D, Williams LE, Wu AM, Yazaki P, Shively JE, Colcher D, Somlo G. A pretherapy biodistribution and dosimetry study of indium-111-radiolabeled trastuzumab in patients with human epidermal growth factor receptor 2-overexpressing breast cancer. Cancer Biother Radiopharm. 2010;25(4):387-94.
  • Disis ML, Wallace DR, Gooley TA, Dang Y, Slota M, Lu H, Coveler AL, Childs JS, Higgins DM, Fintak PA, dela Rosa C, Tietje K, Link J, Waisman J, Salazar LG. Concurrent trastuzumab and HER2/neu-specific vaccination in patients with metastatic breast cancer. J Clin Oncol. 2009;27(28):4685-92.
  • Alvarez-Rueda N, Ladjemi MZ, Béhar G, Corgnac S, Pugnière M, Roquet F, Bascoul-Mollevi C, Baty D, Pèlegrin A, Navarro-Teulon I. A llama single domain anti-idiotypic antibody mimicking HER2 as a vaccine: Immunogenicity and efficacy. Vaccine. 2009;27(35):4826-33.
  • Morse MA, Hobeika A, Osada T, Niedzwiecki D, Marcom PK, Blackwell KL, Anders C, Devi GR, Lyerly HK, Clay TM. Long term disease-free survival and T cell and antibody responses in women with high-risk Her2+ breast cancer following vaccination against Her2. J Transl Med. 2007 Sep 6;5:42.

Product Citations