AUTOMATED AB42 AND TAU ASSAYS ON THE ECIQ UPDATE€¦ · AUTOMATED AB42 AND TAU ASSAYS ON THE ECIQ UPDATE DEVELOPED IN PARTNERSHIP WITH BRISTOL-MYERS SQUIBB AND J&J ORTHO CLINICAL

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