Golimumab (Simponi ®) ELISA Assay Kit

$895.00

The Eagle Biosciences Golimumab (Simponi ®) ELISA Assay Kit is an enzyme immunoassay for the analytical determination of free Golimumab in serum and plasma samples. This assay utilizes the target molecule (TNF-α) as the binding reagent on the microtiter plate so this assay will measure any other therapeutic that targets TNF-α including Adalimumab, Etanercept, and Infliximab.  The Golimumab ELISA Assay Kit is for research use only and not to be used in diagnostic procedures.

Golimumab (Simponi®)ELISA Assay Kit

For Research Use Only

Size: 1 x 96 wells
Sensitivity: 5 ng/mL
Dynamic Range: 6-200 ng/mL
Incubation Time: 2 hours
Sample Type:  Serum, Plasma
Sample Size: 10 µL

Additional Information

Assay Background

Golimumab (Simponi ®) is a human monoclonal antibody that binds to both the soluble and transmembrane bioactive forms of human TNFα. This interaction prevents the binding of TNFα to its receptors, thereby inhibiting the biological activity of TNF. Golimumab has been proven effective in the treatment of Rheumatoid Arthritis (RA), Ankylosing Spondylitis (AS), Psoriatic Arthritis (PsA) or Ulcerative Colitis (UC). Antibodies to Golimumab were detected in 57 (4%) of Golimumab -treated patients across the Phase 3 RA, PsA, and AS trials through Week 24. Similar rates were observed in each of the three indications. Patients who received Golimumab with concomitant Methotrexate (MTX) had a lower proportion of antibodies to Golimumab than patients who received Golimumab without MTX (approximately 2% versus 7%, respectively). Of the patients with a positive antibody response to Golimumab in the Phase 2 and 3 trials, most were determined to have neutralizing antibodies to Golimumab as measured by a cell-based functional assay. The data from the literature demonstrated that Anti-Drug Antibody positivity was significantly associated with low Golimumab levels and poor therapeutic response. The positive correlation between serum drug trough levels and therapeutic response indicates that drug monitoring could be useful for optimising the dosing of biologics in a personalised therapy strategy.

Identification of biomarkers for (non-)response and risk factors for adverse drug reactions that might be related to serum concentrations and maintaining the effective concentration of Golimumab in order to potentially avoid some side effects with a reliable method might be beneficial.

Assay Principle

This Eagle Biosciences Golimumab ELISA Assay Kit is based on sandwich type ELISA. Diluted standards and samples (serum or plasma) are incubated in the microtiter plate coated with recombinant human TNF-α (rh TNF-α). 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 Golimumab pre-captured by the rhTNF-α on the surface of the wells. Following incubation, the wells are washed and the bound enzymatic activity is detected by addition of chromogen-substrate. The color developed is proportional to the amount of free Golimumab in the sample or standard. Results of samples can be determined by using the standard curve.

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 1:50 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 15 min at RT. Avoid exposure to direct sunlight.
  10. Stop the substrate reaction by adding 100 µL of Stop Solution into each well. Briefly mix contents by gently shaking the plate. Color changes from blue to yellow.
  11. 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 naïve serum. Because the solid phase is coated with rhTNF-α, other therapeutic anti-TNF antibodies cause full cross reaction. However, a quantification of other therapeutic antibodies is possible only by using the drug-specific standards, which are available as separate kits from Eagle Biosciences.

Manual

Product Manual


Publications

Citations

  • Gelfer G, Perry L, Deodhar A. Golimumab for the treatment of axial spondyloarthritis. Expert Rev Clin Immunol. 2015; 2:1-10.
  • Lichtenstein GR, Hanauer SB, Sandborn WJ. Emerging Treatment Options in Mild to Moderate Ulcerative Colitis. Gastroenterol Hepatol (N Y). 2015;11(3 Suppl 1):1-16.
  • Emery P, Fleischmann RM, Strusberg I, Durez P, Nash P, Amante E, Churchill M, Park W, Pons-Estel B, Han C, Gathany TA, Xu S, Zhou Y, Leu JH, Hsia EC. Efficacy and safety of subcutaneous golimumab in methotrexate-naïve patients with rheumatoid arthritis: 5-year results of the GO-BEFORE trial. Arthritis Care Res (Hoboken). 2015 Oct 16. doi: 10.1002/acr.22759
  • Lee WK, Kim GW, Cho HH, Kim WJ, Mun JH, Song M, Kim HS, Ko HC, Kim MB, Kim BS. Erythrodermic Psoriasis Treated with Golimumab: A Case Report. Ann Dermatol. 2015 Aug;27(4):446-9.
  • Brady BL, Tkacz JP, Lofland J, Meyer R, Bolge SC. Prescribing Patterns of Intravenous Golimumab for Rheumatoid Arthritis. Clin Ther. 2015;37(9):2028-36.
  • Yonemoto Y, Okamura K, Takeuchi K, Ayabe K, Kaneko T, Matsushita M, Tamura Y, Iso T, Okura C, Otsuka K, Inoue H, Takagishi K. Comparison of golimumab 100-mg monotherapy to golimumab 50 mg plus methotrexate in patients with rheumatoid arthritis: Results from a multicenter, cohort study. Mod Rheumatol. 2015; 3:1-5.
  • Abreu MT Commentary: Golimumab in Moderate-to-Severe Ulcerative Colitis. Gastroenterol Hepatol (N Y). 2014;10(7):455-6.
  • Sandborn WJ, Feagan BG, Marano C, Zhang H, Strauss R, Johanns J, Adedokun OJ, Guzzo C, Colombel JF, Reinisch W, Gibson PR, Collins J, Järnerot G, Hibi T, Rutgeerts P; PURSUIT-SC Study Group. Subcutaneous golimumab induces clinical response and remission in patients with moderate-to-severe ulcerative colitis. Gastroenterology. 2014;146(1):85-95.
  • Thorlund K, Druyts E, Toor K, Mills EJ. Comparative efficacy of golimumab, infliximab, and adalimumab for moderately to severely active ulcerative colitis: a network meta-analysis accounting for differences in trial designs. Expert Rev Gastroenterol Hepatol. 2015;9(5):693-700.
  • Kanbe K, Chiba J, Inoue Y, Taguchi M, Yabuki A. Predictive factors related to the efficacy of golimumab in patients with rheumatoid arthritis. Clin Med Insights Arthritis Musculoskelet Disord. 2015;8:25-32.
  • McCarty DJ. Golimumab intravenous infusion for the treatment of rheumatoid arthritis. Nurse Pract. 2015;40(3):15-8.
  • Smolen JS, Kay J, Doyle M, Landewé R, Matteson EL, Gaylis N, Wollenhaupt J, Murphy FT, Xu S, Zhou Y, Hsia EC. Golimumab in patients with active rheumatoid arthritis after treatment with tumor necrosis factor α inhibitors: findings with up to five years of treatment in the multicenter, randomized, double-blind, placebo-controlled, phase 3 GO-AFTER study. Arthritis Res Ther. 2015 Jan 22;17:14. doi: 10.1186/s13075-015-0516-6.
  • Sato E, Tanaka E, Nakajima A, Inoue E, Shimizu Y, Yamaguchi R, Ochiai M, Shidara K, Hoshi D, Sugimoto N, Seto Y, Taniguchi A, Momohara S, Yamanaka H. Assessment of the effectiveness of golimumab 50-mg and 100-mg regimens in patients with rheumatoid arthritis in daily practice. Mod Rheumatol. 2015;25(4):528-33.
  • Selmi C, Ceribelli A, Naguwa SM, Cantarini L, Shoenfeld Y. Safety issues and concerns of new immunomodulators in rheumatology. Expert Opin Drug Saf. 2015;14(3):389-99.
  • Doyle MK, Rahman MU, Frederick B, Birbara CA, de Vries D, Toedter G, Wu X, Chen D, Ranganath VK, Westerman ME, Furst DE. Effects of subcutaneous and intravenous golimumab on inflammatory biomarkers in patients with rheumatoid arthritis: results of a phase 1, randomized, open-label trial. Rheumatology (Oxford). 2013;52(7):1214-9.
  • Chen DY, Chen YM, Hung WT, Chen HH, Hsieh CW, Chen YH, Huang WN, Hsieh TY. Immunogenicity, drug trough levels and therapeutic response in patients with rheumatoid arthritis or ankylosing spondylitis after 24-week golimumab treatment. Ann Rheum Dis. 2015 Oct 6. pii: annrheumdis-2015-207978. doi: 10.1136/annrheumdis-2015-207978.
  • Huynh C, Reguiai Z, Lambrecht I, Brochot P, Eschard JP, Salmon JH. First two cases of alopecia areata during golimumab therapy. Joint Bone Spine. 2015 Oct 7. pii: S1297-319X(15)00178-5. doi: 10.1016/j.jbspin.2015.08.007. [Epub ahead of print]
  • Brunasso AM, Aberer W, Massone C. Subacute lupus erythematosus during treatment with golimumab for seronegative rheumatoid arthritis. Lupus. 2014;23(2):201-3.
  • Zhuang Y, Lyn S, Lv Y, Xu Z, Bouman-Thio E, Masterson T, Ford JA, Keen M, Petty KJ, Davis HM, Zhou H. Pharmacokinetics and safety of golimumab in healthy Chinese subjects following a single subcutaneous administration in a randomized phase I trial. Clin Drug Investig. 2013;33(11):795-800.
  • Murdaca G, Spanò F, Puppo F. Selective TNF-α inhibitor-induced injection site reactions. Expert Opin Drug Saf. 2013;12(2):187-93.