ADAMTS13 Evaluation

- ADAMTS13 Activity
- ADAMTS13 Antibody Inhibitory Titer

Application: For diagnosis and clinical evaluation of thrombotic thrombocytopenic purpura

(TTP) patients.
Test Features:
- High detection sensitivity of ADAMTS13 activity (range of 0.5-100%)
- Determine functional titer of ADAMTS13 antibody
- Clinical consultation & laboratory interpretation available
Test price: $200
Turnaround time: 2-3 days
Sample Requirements: 0.5mL fresh frozen citrated plasma, shipped on dry ice.
To Order: Institution MUST set up an account with Ohio State University Medical Center
(OSUMC) University Reference Lab. See contact information below.

For technical inquiries:

OSU Coagulation Reference Laboratory TO SET UP AN ACCOUNT,
1645 Neil Avenue – 150, Hamilton Hall Or for ordering & billing questions contact:
Columbus, OH 43210 Ohio State University
p. 614.292.1798 Reference Laboratories (URL)
f. 614.292.3144 1033 North High Street
e-mail: april.horne@osumc.edu Columbus, OH 43201
p. 1.800.227.5062
http://pathology.osumc.edu/ext/research/wulab/CRL.htm f. 614.293.9280

Method illustrations:
ADAMTS13 Activity: SELDI-TOF
mass spectrometer combines the
ProteinChip surface chemistry and time
of flight mass analyzer to detect and
measure the analyte derived from
enzymatic action of ADAMTS13 (A
Disintegrin And Metalloprotease with
ThromboSpondin type 1 repeats). In this
assay, a vWF fusion protein with a
lysine tag is used as substrate for
ADAMTS13. After cleavage, a 7540
Dalton peptide containing a 5X lysine
tag is generated in an amount
corresponding to the plasma
ADAMTS13 activity in TTP patients.
The cleaved product is selected
specifically by CM10 cationic
proteinChip and then measured by
SELDI-TOF mass spectrometer.
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As shown in the figures, the peak of the analyte (7,540 dalton) exhibits an excellent dose response
curve corresponding to the amount of ADAMTS13 activity in the sample. When the quantity of the
analyte is normalized by the internal control
peptide, there is an excellent linear relationship
between analyte readout and amount of
ADAMTS13 calibrators (r2 = 0.999). The test also
offers excellent test reproducibility (CV of less
than 20%). When the test is slightly modified, it
can detect the ADAMSTS13 activity as low as
0.5% reproducibly. This improvement allows for
accurate examination of ADAMTS13 activity levels
in greater than 95% of samples from TTP patients.

60%

50% y = 0.1746x - 2E-05

R2 = 0.9997
% ADAMTS 13 activity

40%

30%

20%

10%

0%
0 0.5 1 1.5 2 2.5 3
Ratio of peak area

ADAMTS13 Antibody Inhibitory Titer: Determination of ADAMTS13 autoantibody inhibitory titers is

performed using a similar approach as described for the determination of Bethesda unit (BU) in patients with a
factor inhibitor. ADAMTS13 activity is determined after pooled normal plasma (PNP) is mixed with an equal
volume of patient plasma made at various dilutions. Afterwards, the ADAMTS13 activity from each sample
gets divided by the ADAMTS13 activity obtained from PNP control to obtain the residual activity. The BU is
then extrapolated from the residual activity according to the standard Bethesda graph. This assay will not be
performed if Activity is reported as normal.
References
1. Cataland S, Jin M, Lin S, Kennedy M, Kraut E, George J and Wu H: Concurrent Cyclosporine and Plasma
Exchange in Idiopathic Thrombotic Thrombocytopenic Purpura. British Journal of Haematology, 139(3):
486-493, 2007.
2. Cataland S, Jin M, Ferketich A, Kennedy K, Kraut E, George J, and Wu H: An Evaluation of Cyclosporine
and Corticosteroids Individually as Adjuncts to Plasma Exchange in the Treatment of Thrombotic
Thrombocytopenic Purpura. British Journal of Haematology, 136 (1) 146-149, 2007.
3. Cataland S and Wu H: Targeting the Inhibitor of ADAMTS13 in Thrombotic Thrombocytopenic Purpura.
Expert Opinion on Pharmacotherapy, 8(4) 437-444, 2007.
4. Wu H, Cataland S, Bissell M and Jin M: Application of SELDI–TOF Mass Spectrometry in Clinical
Evaluation of Thrombotic Thrombocytopenic Purpura. Spectroscopy 20 (5-6), 219-227, 2007.
5. Cataland S, Jin M, Smith E, Stanek M, and Wu H: Full Evaluation of an Acquired Case of TTP Following
the Surgical Resection of Glioblastoma Multiforme. Journal of Thrombosis Haemostasis. 4(12): 2733-7,
2006.
6. Cataland S, Jin M, Zheng L, George J, and Wu H: An Evaluation of Cyclosporine Alone For the Treatment
of Early Recurrences of TTP. Journal of Thrombosis and Hemostasis, 4, 1162-1164, 2006
7. Jin M, Cataland S, Bissell M, and Wu H: A rapid test for the diagnosis of Thrombotic Thrombocytopenic
Purpura using surface enhanced laser desorption and ionization time-of-flight mass spectrometry. Journal
of Thrombosis and Hemostasis, 4: 333–338. 2006
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