Z Gastroenterol 2017; 55(08): e57-e299
DOI: 10.1055/s-0037-1604810
Kurzvorträge
Dünndarm und Dickdarm, Proktologie
CED: Grundlagen und Biomarker: Donnerstag, 14 September 2017, 08:00 – 09:28, Florenz/Forschungsforum 3
Georg Thieme Verlag KG Stuttgart · New York

An enzyme-linked immunosorbent assay for therapeutic drug monitoring of Golimumab

C Berger
1   Immundiagnostik AG, Bensheim, Deutschland
,
A Pilch
1   Immundiagnostik AG, Bensheim, Deutschland
,
J Ruppert
1   Immundiagnostik AG, Bensheim, Deutschland
,
FP Armbruster
1   Immundiagnostik AG, Bensheim, Deutschland
,
J Stein
2   Interdisciplinary Crohn-Colitis Centrum Rhein-Main, Frankfurt/Main, Deutschland
3   DGD Clinics Sachsenhausen, Dept. of Gastroenterology and Clinical Nutrition, Frankfurt am Main, Deutschland
› Author Affiliations
Further Information

Publication History

Publication Date:
02 August 2017 (online)

 

Introduction:

Golimumab is a therapeutic anti-TNF monoclonal antibody approved for use in moderate to severe ulcerative colitis (UC). The PURSUIT trials showed a significant exposure-response relationship of golimumab in UC. Interindividual differences in response to golimumab treatment may be explained in part by interindividual variability in pharmacokinetics.

Aim:

The aim was to develop and validate an enzyme-linked immunosorbent assay (ELISA) for the detection of golimumab drug concentration.

Methods:

Microtiter plates were coated with recombinant human TNF-alpha. Samples diluted at 1:100 were added to the microtiter plate for binding, and bound golimumab was detected using mouse anti-human immunoglobulin G1 (HRP-anti h IgG1). Performance characteristics of the assay were determined according to the European in-vitro diagnostic devices directive 98/79/EC.

Results:

The limit of detection (LoD) for golimumab determination in human serum samples was 3.56 ng/mL. The measuring range of the assay was determined to be 0.415 – 22.5 µg/mL. Intra-assay variation (n = 19) was ≤6.6%, while inter-assay variation (n = 11) was ≤5.3%. Linearity testing was performed by analysis of three serially-diluted samples spiked with golimumab; golimumab concentrations measured by the new assay were within 98 – 129% of the expected concentrations. The assay detected no false-positive signals from samples taken from untreated patients. Due to the use of TNF-alpha as a capture reagent, other TNF-alpha blockers were detected.

Conclusion:

This newly-developed ELISA method is rapid, accurate and reproducible. The use of monoclonal antibodies to golimumab could improve the specificity of the assay. The ELISA may be useful not only for pharmacokinetic/pharmacodynamic studies, but also in therapeutic drug monitoring of golimumab.