Cross-Platform Comparison of IL-6 Immunoassays. Maribeth Raines, PhD. Vice President of Laboratory Services. May 17, 2016 Biomarker World Congress
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1 Crss-Platfrm Cmparisn f IL-6 Immunassays Maribeth Raines, PhD Vice President f Labratry Services May 17, 2016 Bimarker Wrld Cngress
2 2016 Bimarker Wrld Cngress 2 Cmpany Overview Funded in 1989 as spin ff frm University f Washingtn Lipid Research Center Ex-Member f CDC-CRMLN (Chlesterl Reference Methd Labratry Netwrk) Relatinships and cllabratins with numerus clinical diagnstic manufacturers CAP CLIA-certified facilities and prcesses but als perate t GLP and bianalytical standards Menu f ver 300 validated assays Platfrm agnstic Established reputatin fr scientific expertise Scientific versight thrughut, frm pre-analytical input t data delivery
3 2016 Bimarker Wrld Cngress 3 Study Objectives Evaluate Cmmercial IL-6 immunassays Use serum samples frm nrmal healthy dnrs and inflammatry disease patients. Parallelism Additinal Perfrmance Parameters Spike and Recvery Functinal Sensitivity Ciraplex Ultra IL-6 US perfrmance QC perfrmance Clinical Trials
4 2016 Bimarker Wrld Cngress 4 IL-6 Cmmercial Immunassay Kits Selectin f the best immunassays depends n the intended use and individual requirements Assay Parameter R&D Systems hsil-6 ELISA Ciraplex ULTRA IL-6 US MSD IL-6 V-plex Erenna IL-6 Kit Price $ $ $ $$$ Length f Test 5 hrs 3.5 hrs 4.5 hrs hrs Assay Frmat ELISA Chemiluminescent Electrchemiluminescent Flurescent Micrparticle Analytical measureable range pg/ml pg/ml pg/ml pg/ml # f Standards Sample vlume (per well) 100 ml 50 ml 50 ml 75 ml MRD NA 1:2 1:2 NA Sensitivity/LLOQ (assigned) 0.08 pg/ml pg/ml 1.58 pg/ml 0.08 pg/ml Nrmal Levels pg/ml < 4.5 pg/ml pg/ml < 5.4 pg/ml
5 2016 Bimarker Wrld Cngress 5 Platfrms fr IL-6 Immunassay Evaluatin Aushn Cira - Brad RQ (3+ Lg) - Multiplexing - Explratry analysis - Fit-fr-purpse - Open platfrm MSD QuickPlex - Well-established - Versatile menu - Multiplexing - Large-scale screening Singulex Erenna - Ultra-lw cncentratin - PK/PD fr bilgics - Prprietary assay transfer - Open platfrm
6 2016 Bimarker Wrld Cngress 6 Standard Curves and Quality Cntrls Ciraplex ULTRA US IL-6 High Cntrl abve Erenna ULOQ and Lw cntrl belw MSD LLOQ IL-6 Assay Standards PBI in-huse IL-6 Matrix Cntrls STD Aushn MSD Erenna (pg/ml) (pg/ml) (pg/ml) QC level QC Lw QC Med QC High Mean (pg/ml) Aushn %CV Mean (pg/ml) MSD %CV Mean (pg/ml) Erenna %CV ~ LLOQ regin
7 2016 Bimarker Wrld Cngress 7 Elevated IL-6 values in 5 f 16 inflammatry samples UC = ulcerative clitis CR = Crhn s Disease PS = Psriasis RA = Rheumatid Arthritis Sample ID Aushn Erenna MSD (pg/ml) (pg/ml) (pg/ml) UC UC UC CR CR CR PS PS PS RA RA RA Sepsis Sepsis Pl > Sepsis Pl Sepsis Pl > NHS NHS NHS NHS n Distributin f Ciraplex ULTRA IL-6 levels in Serum Samples < >200 Range (pg/ml) QCL = 0.25 QCM = 6.50 QCM = Inflammatry Samples NHS Samples >ULOQ %CV >20% <LLOQ
8 2016 Bimarker Wrld Cngress 8 Platfrm Crrelatin Analysis - Ttal Better crrelatin with MSD ; better value agreement with Erenna althugh sme values > Erenna ULOQ
9 2016 Bimarker Wrld Cngress 9 Platfrm Crrelatin Analysis (< 10 pg/ml) MSD and Erenna crrelate with each ther and have 25-40% lwer values than Ciraplex ULTRA
10 2016 Bimarker Wrld Cngress 10 All three platfrms exhibit gd Parallelism t the LLOQ Diseased Serum Samples Nrmal Serum Samples Arrw indicates value nearest LLOQ
11 2016 Bimarker Wrld Cngress 11 Functinal Sensitivity LLOQ Cmparisn Ciraplex ULTRA IL-6 US exhibits best IL-6 sensitivity MSD V-plex (IL-6 pg/ml) Sample Rep 1 Rep 2 Rep 3 Mean %CV LLOQ LLOQ Ciraplex Ultra IL-6 US Graph LLOQ LLOQ LLOQ LLOQ LLOQ = pg/ml Erenna IL-6 (pg/ml) Sample Rep 1 Rep 2 Rep 3 Mean %CV LLOQ LLOQ LLOQ LLOQ LLOQ LLOQ LLOQ = pg/ml Ciraplex ULTRA IL-6 US (pg/ml) Sample Rep 1 Rep 2 Rep 3 Mean %CV LLOQ LLOQ LLOQ LLOQ LLOQ LLOQ LLOQ = pg/ml
12 2016 Bimarker Wrld Cngress 12 Platfrm Cmparisn Summary Alternative t classic R&D systems ELISA? Ciraplex ULTRA and MSD V-plex cver several lgs f IL-6 values Erenna issues with duplicate CV failures, lw thrughput, and high kit csts makes it a less attractive Ciraplex ULTRA IL-6 US values Parallelism 40 and 60% higher than MSD V-plex when IL-6 > 100 pg/ml and < 10 pg/ml, respectively 25% higher than Erenna platfrm Parallelism acceptable t LLOQ fr NHS and inflammatry samples fr all platfrms Ciraplex ULTRA parallelism at 1:4 MRD is better than 1:2 Accuracy Spike Recvery Sensitivity Recvery better at lw cncentratins fr Ciraplex ULTRA while Erenna and MSD V-plex yields better recveries at higher cncentratins Ciraplex ULTRA IL-6 US has best sensitivity with pg/ml MSD LLOQ better than listed in kit insert;clse t ~ 0.1 pg/ml, but belw Ciraplex lw QC
13 2016 Bimarker Wrld Cngress 13 Ciraplex ULTRA IL-6 Perfrmance in Clinical Studies Ciraplex ULTRA IL-6 US perfrmance range reduces repeat rate fr clinical trial studies Clinical Studies Over 2500 samples Cllected ver a year span As many as 24 plates (~750 samples) analyzed per day IL-6 bserved values Range frm pg/ml Detects changes in patients ver 2-3 lgs > ULOQ (228 pg/ml); N = 1 < LLOQ (0.016 pg/ml); N = 2 > 10 pg/ml; N = 324 Mean IL-6 value = 4.832; Median IL-6 values = 2.848
14 SD 2016 Bimarker Wrld Cngress 14 Cnsistent Perfrmance f the Ciraplex ULTRA IL-6 US ver time and acrss clinical studies Run Date
15 2016 Bimarker Wrld Cngress 15 Ciraplex ULTRA IL-6 US is the right chice fr clinical trial studies Practical Cnsideratins Price is right Shrt incubatin times Small sample vlume Amenable t autmatin, s high thrughput Perfrmance Excellent sensitivity - able t detect 100% nrmal samples and levels belw nrmal Range is high enugh t detect mst inflammatry samples except maybe sepsis patients. Can detect changes in IL-6 values in patients ver several lgs Recvery is gd where it cunts; at the lw end 1:4 MRD may imprve perfrmance even mre Kits d nt include assay cntrls which may be an issue fr sme labs
16 2016 Bimarker Wrld Cngress 16 Acknwledgements PBI Sarah Christensen Erin Brwn Lenard Egglestn Tnya Aggune Amar Sethi A special thanks t Aushn Bisystems fr their spnsrship f this prject.
17 2016 Bimarker Wrld Cngress 17 Any Questins?
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