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TOCILIZUMAB ELISA (MAB-BASED)
Enzyme immunoassay for the specific and quantitative determination of free Tocilizumab (Actemra®) in serum and plasma.
The solid phase (MTP) is coated by a highly specific monoclonal antibody directed against Tocilizumab. Another POD-labelled specific monoclonal antibody is used as conjugate.
Therefore any cross reactivity to the other therapeutical monoclonal antibodies is excluded.
| Required Volume (µL) | 10 |
| Incubation Time (min) | 130 |
| Sample | Serum or Plasma |
| Plate Size | 96 |
| 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 and diagnostic uses. It is suitable for Therapeutical Drug Monitoring (TDM) purposes.
ESSAY CHARACTERISTICS
SPECIFICITY
There is no cross reaction with any other proteins present in native human serum. A screening test was performed with 32 different native human sera. All produced OD450/620nm 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 Adalimumab, Golimumab, Infliximab, Etanercept, Rituximab, Bevacizumab, Trastuzumab and Omalizumab at concentrations up to 1 mg/mL. All produced OD450/620nm values less than the mean OD of standard D.
SENSITIVITY
The lowest detectable level that can be clearly distinguished from the zero standard is 2 ng/mL (zero standard +2SD read from the curve) under the above-described conditions. Analytical sensitivity is 2 ng/mL, and corresponding to the detection limit (limit of quantification) of 2 µg/mL for undiluted clinical samples because the serum or plasma samples are instructed to be diluted at 1:1000 before starting the assay.
PRECISION OF THE KIT
Intra-assay CV: <10%.
Inter-assay CV: <10%.
RECOVERY
Recovery rate was found to be >95% with native human serum and plasma samples when spiked with exogenous Tocilizumab.
AUTOMATION
The ImmunoGuide Tocilizumab ELISA (mAb-based) is suitable also for being used by an automated ELISA processor.
REFERENCES
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2. Kennedy GA, Varelias A, Vuckovic S, Le Texier L, Gartlan KH, Zhang P, Thomas G, Anderson L, Boyle G, Cloonan N, Leach J, Sturgeon E, Avery J, Olver SD, Lor M, Misra AK, Hutchins C, Morton AJ, Durrant ST, Subramoniapillai E, Butler JP, Curley CI, MacDonald KP, Tey SK, Hill GR. Addition of interleukin-6 inhibition with tocilizumab to standard graft-versus-host disease prophylaxis after allogeneic stem-cell transplantation: a phase 1/2 trial. Lancet Oncol. 2014;15(13):1451-9
3. Besada E. Potential patient benefit of a subcutaneous formulation of tocilizumab for the treatment of rheumatoid arthritis: a critical review. Patient Prefer Adherence. 2014;8:1051-9.
4. Ohta S, Tsuru T, Terao K, Mogi S, Suzaki M, Shono E, Ishida Y, Tarumi E, Imai M. Mechanism-based approach using a biomarker response to evaluate tocilizumab subcutaneous injection in patients with rheumatoid arthritis with an inadequate response to synthetic DMARDs (MATSURI study). J Clin Pharmacol. 2014;54(1):109-19.
5. Lee SJ, Park W, Park SH, Shim SC, Baek HJ, Yoo DH, Kim HA, Lee SK, Leee YJ, Park YE, Cha HS, Park JK, Lee EY, Lee EB, Song YW. Low baseline interleukin-17A levels are associated with better treatment response at 12 weeks to tocilizumab therapy in rheumatoid arthritis patients. J Immunol Res. 2015;2015:487230. doi: 10.1155/2015/487230. Epub 2015 Apr 2
6. Ogata A, Tanimura K, Sugimoto T, Inoue H, Urata Y, Matsubara T, Kondo M, Ueki Y, Iwahashi M, Tohma S, Ohta S, Saeki Y, Tanaka T; Musashi Study Investigators. Phase III study of the efficacy and safety of subcutaneous versus intravenous tocilizumab monotherapy in patients with rheumatoid arthritis. Arthritis Care Res (Hoboken). 2014;66(3):344-54.
7. Mori S, Ueki Y. Primary lack of efficacy of infliximab therapy for rheumatoid arthritis: pharmacokinetic characterization and assessment of switching to tocilizumab. Mod Rheumatol. 2011;21(6):628-36.
8. Zhang X, Peck R. Clinical pharmacology of tocilizumab for the treatment of patients with rheumatoid arthritis. Expert Rev Clin Pharmacol. 2011 Sep;4(5):539-58. doi: 10.1586/ecp.11.33.
9. Burmester GR, Rigby WF, van Vollenhoven RF, Kay J, Rubbert-Roth A, Kelman A, Dimonaco S, Mitchell N. Tocilizumab in early progressive rheumatoid arthritis: FUNCTION, a randomised controlled trial. Ann Rheum Dis. 2015 Oct 28. pii: annrheumdis-2015-207628. doi: 10.1136/annrheumdis-2015-207628. [Epub ahead of print]
10. Gabay C, Riek M, Hetland ML, Hauge EM, Pavelka K, Tomšič M, Canhao H, Chatzdionysiou K, Lukina G, Nordström DC, Lie E, Ancuta I, Hernández MV, van Riel PL, van Vollenhoven R, Kvien TK. Effectiveness of tocilizumab with and without synthetic disease-modifying antirheumatic drugs in rheumatoid arthritis: results from a European collaborative study. Ann Rheum Dis. 2015 Sep 15. pii: annrheumdis-2015-207760. doi: 10.1136/annrheumdis-2015-207760
11. Espinoza F, Jorgensen C, Pers YM. Efficacy of Tocilizumab in the treatment of Eosinophilic fasciitis: Report of one case. Joint Bone Spine. 2015 Jul 7. pii: S1297-319X(15)00101-3. doi: 10.1016/j.jbspin.2015.02.008. [Epub ahead of print]
12. Roddy JV, Haverkos BM, McBride A, Leininger KM, Jaglowski S, Penza S, Klisovic R, Blum W, Vasu S, Hofmeister CC, Benson DM, Andritsos LA, Devine SM, Efebera YA. Tocilizumab for steroid refractory acute graft-versus-host disease. Leuk Lymphoma. 2015 Jul 3:1-5. [Epub ahead of print] 9.
13.Kim NH, Kim SK, Kim DS, Zhang D, Park JA, Yi H, Kim JS, Shin HC. Anti-proliferative action of IL-6R-targeted antibody tocilizumab for non-small cell lung cancer cells. Oncol Lett. 2015;9(5):2283-2288.
14. Hammoudeh M, Al Awadhi A, Hasan EH, Akhlaghi M, Ahmadzadeh A, Sadeghi Abdollahi B Safety, Tolerability, and Efficacy of Tocilizumab in Rheumatoid Arthritis: An Open-Label Phase 4 Study in Patients from the Middle East. Int J Rheumatol. 2015;2015:975028. doi: 10.1155/2015/975028.
15. Yang L, Xing R, Li C, Liu Y, Sun L, Liu X, Wang Y. Active immunization with Tocilizumab mimotopes induces specific immune responses. BMC Biotechnol. 2015 Jun 2;15:46. doi: 10.1186/s12896-015-0161-9.
16. Burmester GR, Rubbert-Roth A, Cantagrel A, Hall S, Leszczynski P, Feldman D, Rangaraj MJ, Roane G, Ludivico C, Bao M, Rowell L, Davies C, Mysler EF. Efficacy and safety of subcutaneous tocilizumab versus intravenous tocilizumab in combination with traditional DMARDs in patients with RA at week 97 (SUMMACTA). Ann Rheum Dis. 2015 Jun 8. pii: annrheumdis-2015-207281. doi: 10.1136/annrheumdis-2015-207281.
17. Romão VC, Santos MJ, Polido-Pereira J, Duarte C, Nero P, Miguel C, Costa JA, Bernardes M, Pimentel-Santos FM, Barcelos F, Costa L, Melo Gomes JA, Pereira da Silva JA, Cunha Branco J, Canas da Silva J, Pereira da Silva JA, Fonseca JE, Canhão H. Comparative Effectiveness of Tocilizumab and TNF Inhibitors in Rheumatoid Arthritis Patients: Data from the Rheumatic Diseases Portuguese Register, Reuma.pt. Biomed Res Int. 2015;2015:279890. doi: 10.1155/2015/279890
18. Wang G, Mu R, Xu H. Management of rheumatoid arthritis in People's Republic of China - focus on tocilizumab and patient considerations. Int J Gen Med. 2015;8:187-94.
19. Bensaoud N, Rostom S, Bahiri R, Hajjaj-Hassouni N. Efficacy of tocilizumab on MRI-determined bone oedema in rheumatoid arthritis. Clin Rheumatol. 2015;34(6):1031-7.
20. Narváez J, Díaz-Torné C, Magallares B, Hernández MV, Reina D, Corominas H, Sanmartí R, de la Serna AR, Llobet JM, Nolla JM. Comparative effectiveness of tocilizumab with either methotrexate or leflunomide in the treatment of rheumatoid arthritis. PLoS One. 2015 Apr 1;10(4):e0123392. doi: 10.1371/journal.pone.0123392. eCollection 2015.
INSTRUCTIONS FOR USE
SAFETY DATA SHEET
BATCH/LOT INFORMATION