AUTOMATED AB42 AND TAU
ASSAYS ON THE ECIQ UPDATE
DEVELOPED IN PARTNERSHIP WITH
BRISTOL-MYERS SQUIBB AND J&J ORTHO CLINICAL DIAGNOSTICS
Salvatore J. Salamone July 14, 2012
Vancouver
DIAGNOSTIC PLATFORM VITROS® ECIQ
IMMUNODIAGNOSTIC SYSTEM
• 30 reagent channels • 90 samples/hour • Sample types: Serum, Plasma, Urine, Amniotic
Fluid, Whole Blood
• Sample volume: 10-80μL • Sample capacity: 60 samples • Disposable tip metering verifies sample
aspiration and dispense and addresses carryover and cross-contamination concerns
• Clot, bubble, low and high viscosity, thin layer fluid and short sample
• Sample trays accommodate: •10mL, 7mL, 5mL collection tubes •1.5mL micro-collection containers • VITROS Microsample cups and 0.5mL and 2.0mL cups
Advantages: • Short analysis time (amenable to quick turn-around – e.g. 1 hr) • Minimal requirements for lab tech time • Robust and precise assays • FDA-cleared platform
CSF Ab42 ASSAY Update
STANDARD CURVE FOR CALIBRATION OF Aβ42
0
10000
20000
30000
40000
50000
60000
0 500 1000 1500 2000
ALU
Aβ42 pg/mL
Test Range ±2SD on control ALUs
0
500
1000
1500
2000
2500
3000
3500
4000
4500
5000
0 100 200 300
ALU
Aβ42 pg/mL
Lower Concentrations ±2SD on control ALUs
Ab42 REAGENT STABILITY
• The proposed reagent formulations are stable at 2-8 oC for up to 7 months
• Stability studies are ongoing
80
90
100
110
120
130
140
0 4 8 12 16 20 24 28 32
AB
42
D
ose
(n
g/m
L)
Week
AB-42 Reagent Stability
QCB (2-8 C)
Mean 2-8 Data
3SD Limit*
450
500
550
600
650
0 4 8 12 16 20 24 28 32
AB
42
D
ose
(n
g/m
L)
Week
AB-42 Reagent Stability
QCD (2-8 C)
Mean 2-8 Data
3SD Limit*
SYNTHETIC Aβ42 CALIBRATOR VS. NATIVE Aβ42
25
250
2,500
25,000
25 250 2,500.
Native Peptide #3
Log1
0 Si
gnal
(ALU
)
Log10 [Analyte] (pg/ml)
Ab42 CALIBRATOR STABILITY
• The proposed calibrator formulation is stable at 2-8 oC for up to 39 weeks
• Stability studies are ongoing
-25.0
-20.0
-15.0
-10.0
-5.0
0.0
5.0
10.0
15.0
20.0
25.0
0 5 10 15 20 25 30 35 40 45
% D
iffe
ren
ce f
ro
m F
ro
zen
Time (Weeks)
VITROS AB-42Calibrator Stability (Liquid 2-8 C vs Frozen -20 C))
Cal 3 Cal 2
Ab42 PRECISION PERFORMANCE
5-Day Precision Study – 4 individual CSF Samples and 3 Controls % CV pg/mL, n=20 (4 reps/day)
CSF129 113 4.89 4.3 4.1 1.7
CSF220 847 28.7 3.4 3.2 1.1
CSF239 422 15.0 3.6 3.4 1.5
CSF278 244 14.9 6.1 5.8 1.5
QCB (R&D) 121 0.94 0.8 0.7 0.5
QCD (R&D) 614 5.07 0.8 0.8 0.6
QCE (R&D) 1167 9.27 0.8 0.8 0.5
CSF Mean 4.1 1.4
QC Mean 0.8 0.5
AB-42 Reagent with Hinged BJ/c
Sample ID Mean SD %CVWithin-lab
Precision %CV
Repeatability
%CV
VITROS AB42 ASSAY LINEARITY
AB42 ASSAY: HIGH DOSE HOOK
No high dose hook up to ~40,000 pg/mL
TAU ASSAY
CSF Tau Assay Technical Performance
STANDARD CURVE FOR CALIBRATION OF TAU
0
1000
2000
3000
4000
5000
6000
7000
8000
9000
0 1000 2000 3000 4000
ALU
Tau pg/mL
Test Range ±2SD on control ALUs
0
200
400
600
800
1000
1200
0 100 200 300 400 500
ALU
Tau pg/mL
Lower Concentrations ±2SD on control ALUs
TAU LIQUID CALIBRATOR STABILITY
• The proposed calibrator formulation is stable at 2-8 oC for up to 90 days
• Stability studies are on-going
50%
60%
70%
80%
90%
100%
110%
120%
130%
140%
150%
0 20 40 60 80 100
% R
ecov
ery
Time (Days)
Peptide Calibrator Stability At 4C
Low Cal
High Cal
TAU PRECISION PERFORMANCE
0.0
1.0
2.0
3.0
4.0
5.0
% C
V
Tau Assay Precision (% CV) of Buffer Controls and Pooled CSF n=4
Low Control (207pg/mL)
Med Control (766pg/mL)
High Control (3150pg/mL)
Low CSF Pool (459pg/mL)
Med CSF Pool (599pg/mL)
High CSF Pool (826pg/mL)
TAU CSF DILUTION LINEARITY
0
20
40
60
80
100
120
2X 4X
% R
ecov
ery
CSF Dilution Factor
Dilution Of Four Individual CSF's Using OCD High Sample Diluent B
CSF 1
CSF 2
CSF 3
CSF 4
TAU ASSAY: HIGH DOSE HOOK
0
50000
100000
150000
200000
250000
300000
350000
400000
0 20000 40000 60000 80000 100000
Ligh
t U
nit
s
Ref Cal Conc (pg/mL)
Tau Assay High Dose Hook Curve
No high dose hook up to ~100,000pg/mL
UPDATE SUMMARY
• Both assys are in Design Lock
• Assays exhibit linearity upon dilution
• No evidence of a “hook effect”
• Calibrators and reagents show good
real time stability at 4°C
• Reagents and calibrators are in process
development at production facility
• Poster at this meeting on Wednesday:
P4-346
ACKNOWLEDGEMENTS
• BMS
– George Green
– Holly Soares
– Paul Rhyne
– Flora Berisha
– RJ Neely
• Ortho Clinical Diagnostics
– Paul Contestable
– Shari Jackson
– Karen Ackles
– Anne Tweedie
– Deb Byrne
– Carmen Vullo
– Ellen Oliver
– Lisa DiMagno
– Susan Werner
– Stacey Alvey
– Lucius Fox
• Saladax
– Irina Baburina
– Gary Feiss
– Jodi Courtney
– Chris Spedaliere
– Hadia Riaz
– Caitlin Baldasano
– Alex Volkov
– Kim Williams
– Dan Kozo
– Rebecca Harney
– Suzanne Pruzinsky
– Michael Lacerda
• AJ Simon Enterprises
– Adam J Simon