Test Code ECMP Eculizumab Monitoring Panel, Serum
Ordering Guidance
To measure only serum concentration of eculizumab, order ECULI / Eculizumab, Serum.
Specimen Required
Patient Preparation:
1. Fasting preferred.
2. Suggest discontinuing natalizumab at least 4 weeks prior to testing for eculizumab quantitation in serum. Patient should consult the healthcare provider who prescribed this drug to determine if discontinuation is an option. If not, ok to proceed with testing while taking natalizumab.
Supplies: Sarstedt Aliquot Tube, 5 mL (T914)
Collection Container/Tube: Red top (serum gel/SST are not acceptable)
Submission Container/Tube: 2 Plastic vials
Specimen Volume: 2 mL in 2 plastic vials, each vial containing 1 mL
Collection Instructions:
1. Draw blood immediately before next scheduled dose.
2. Immediately after specimen collection, place the tube on wet ice.
3. After sample has clotted on wet ice, centrifuge at 4° C and aliquot serum into two 5 mL plastic vials.
4. Freeze specimen within 30 minutes of centrifugation. Sample must be placed on dry ice if not frozen immediately.
Secondary ID
618696Useful For
Monitoring of complement blockage by eculizumab
Assessing the response to eculizumab therapy
Assessing the need for dose escalation
Evaluating the potential for dose de-escalation or discontinuation of therapy in remission states
Monitoring patients who need to be above a certain eculizumab concentration to improve the odds of a clinical response for therapy optimization
This test is not useful as the sole basis for a diagnosis or treatment decisions.
Profile Information
Test ID | Reporting Name | Available Separately | Always Performed |
---|---|---|---|
ECULI | Eculizumab, S | Yes | Yes |
EAH50 | Eculizumab Complement Blockage, S | No | Yes |
ECUIN | Eculizumab Interpretation, S | No | Yes |
Method Name
EAH50: Enzyme-Linked Immunosorbent Assay (ELISA)
ECULI: Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS), High Resolution Accurate Mass
Specimen Type
SerumSerum Red
Specimen Minimum Volume
1 mL total in 2 plastic vials, each vial containing 0.5 mL
Specimen Stability Information
Specimen Type | Temperature | Time | Special Container |
---|---|---|---|
Serum | Frozen | 14 days | |
Serum Red | Frozen | 14 days |
Reject Due To
Gross hemolysis | Reject |
Gross lipemia | Reject |
Gross icterus | OK |
Reference Values
Eculizumab Complement Blockage:
≥46% normal
Eculizumab:
Lower limit of quantitation =5.0 mcg/mL
>35 Therapeutic concentration for paroxysmal nocturnal hemoglobinuria (PNH)
>50 Therapeutic concentration for atypical hemolytic uremic syndrome (aHUS)
Interpretation
Minimum trough therapeutic concentrations (immediately before next infusion) of eculizumab are expected to be above 35 mcg/mL for paroxysmal nocturnal hemoglobinuria and above 50 mcg/mL for atypical hemolytic uremic syndrome.
For the complement blockage monitoring of eculizumab:
-When eculizumab is present in serum at concentrations around 100 mcg/mL, the results are below the limit of quantitation of the assay (<10% of normal).
Method Description
The Wieslab enzyme-linked immunosorbent assay (ELISA) complement assay for the alternative pathway combines principles of the hemolytic assay for complement activation with the use of labeled antibodies specific for neoantigens produced as a result of complement activation. The microtiter plate strips are coated with lipopolysaccharide. Patient serum is diluted in diluent containing specific blocker to ensure that only the alternative pathway is activated. During the first incubation, the diluted patient serum in the wells is activated by the coating. The wells are then washed and C5b-9 (membrane attack complex: MAC) is detected with a specific alkaline phosphatase labeled antibody to the neoantigen expressed during MAC formation. After a final wash, an alkaline phosphatase substrate is added. The amount of alternative pathway complement activity correlates with the color intensity of the solution and is measured in terms of absorbance (optical density).(Nordin JG, Truedsson L, Sjoholm A. New procedure for detection of complement deficiency by ELISA. Analysis of activation pathways and circumvention of rheumatoid factor influence. J Immunol Methods. 1993;166(2):263-270; Frazer-Abel A, Sepiashvili L, Mbughuni MM, Willrich MA. Overview of laboratory testing and clinical presentations of complement deficiencies and dysregulation. Adv Clin Chem. 2016;77:1-75. doi:10.1016/bs.acc.2016.06.001)
Eculizumab is extracted from serum and measured by liquid chromatography (high-resolution accurate-mass) mass spectrometry.(Unpublished Mayo method)
Day(s) Performed
Varies
Report Available
3 to 12 daysSpecimen Retention Time
14 daysPerforming Laboratory
Mayo Clinic Laboratories in RochesterCPT Code Information
80299
86161
LOINC Code Information
Test ID | Test Order Name | Order LOINC Value |
---|---|---|
ECMP | Eculizumab Monitoring Panel, S | 101922-3 |
Result ID | Test Result Name | Result LOINC Value |
---|---|---|
65676 | Eculizumab, S | 90240-3 |
619951 | Eculizumab Interpretation | 59462-2 |
618697 | Eculizumab Complement Blockage, S | 74520-8 |
Forms
If not ordering electronically, complete, print, and send a Coagulation Test Request (T753) with the specimen.