TECHNOZYM® ADAMTS13
Chromogenic and fluorogenic ELISAs for determination of ADAMTS13 antigen and inhibitor concentration as well as ADAMTS13 activity in human plasma
Your partner for ADAMTS13 TECHNOZYM® ADAMTS13
A disintegrin and metalloprotease with thrombospondin motifs
ADAMTS13 is a plasma protein with low concentration (5 nmol/L) which cleaves very specific large vWF multimeres under laminar flow conditions A functional defect of ADAMTS13 leads to presence of higher molecular weight forms of vWF in plasma and thus to increased platelet aggregation → Main cause for thrombotic thrombocytopenic purpura (TTP) Thrombotic microangiopathy (TMA) is a pathologic state which results in thrombosis in capillaries and arterioles, due to an endothelial injury. The classic TMAs are aquired hemolytic uremic syndro- me (aHUS) and TTP:
≤ 5 % ADAMTS-Activity ≥ 5 % ADAMTS-Activity TTP aHUS
ADAMTS13 FUNCTION according to Tsai H-M. Int. J. Hematol. 2010;91:1-9
Intact vessel wall ADAMTS13 regulates under normal circumstances the size of vWF multimers
Injured/inflamed vessel wall vWF multimers build a connection between collagen and platelets
Absence of ADAMTS13 Formation of very large vWF multimers leads to platelet aggregation in healthy vessels TTP
Presentation Dr. Scheiflinger, Laibach June 2004 ® KORRELATIONENTECHNOZYM DER ADAMTS13 ADAMTS-13 ANTIGEN AKTIVITÄTSME (chromogenic)SSUNGEN Chromogenic ELISA for determination of ADAMTS13 antigen concentration
Measuring range 0.0 - 1.0 U/ml LOD 0.012 U/ml High linearity Calibrators and controls are calibrated against the WHO standard for ADAMTS13 Fast result within approximately 3.5 hours Detection is independent of enzyme activity Manual or fully automated testing
® TECHNOZYM ADAMTS13 INHIBITOR (chromogenic) Chromogenic ELISA for determination of antibodies directed against ADAMTS13
Differentiation between TTP and other auto antibodies Measuring range 2 - 104 U/ml LOD 1.68 U/ml High specifity Fast result Manual or fully automated testing
TECHNOZYM® ADAMTS13 INH ELISA can differentiate between acute TTP 45 40 caused by anti- ADAMTS13 antibody and 35 patients who are in remission or have 30 other forms of TTP. 25 20
ADAMTS13 ADAMTS13 titer 15 - It is highly specific and can clearly diffe- 10 rentiate between TTP samples and other anti 5 samples with auto-antibodies (e.g. Lupus) 0 or high IgG concentration (e.g. IgG Gammophathy) ® TECHNOZYM ADAMTS13 ACTIVITY (chromogenic) Chromogenic ELISA for the determination of ADAMTS13 activity
Calibration curve High sensitivity for ADAMTS-13 activity down to 2 0.003 IU/ml 1,5 Short assay time (3 h)
1 Calibrators and controls are calibrated against the WHO standard for ADAMTS13 OD OD 450 nm 0,5 No interference by bilirubin, hemolytic or lipemic patient 0 samples 0 20 40 60 80 100 High precision in the decision range with intra-assay ADAMTS-13 Activity [%] CVs <5% and inter-assay CVs <9% Can be used for ADAMTS13 Inhibitor determination in Bethesda assay Demonstration video available on our TECHNOZOOM Youtube Channel (see QR-Code on the last page) TEST PRINCIPLE
CORRELATION
Good correlation in ADAMTS13 activity of different groups of samples ® TECHNOZYM ADAMTS13 (fluorogenic) Fluorogenic ELISA for the determination of ADAMTS13 antigen and activity
TEST PRINCIPLE
ACTIVITY AND ANTIGEN DETERMINATION
Fluorogenic substrate Incubation at room temperature 180 Normals, n=38 TTP patients, n=43 Calculation tool available for antigen and activity 160 Median value: 75 % Median value: 20 % 140 Applications available for various 120 fluorogenic readers 100 80 No sample predillution 60 Calibrators and controls are calibrated against 40
20 the WHO standard for ADAMTS13
0
ADAMTS-13 activity in % in activity ADAMTS-13 -20 Normals, n=38 300 TTP patients, n=43 -40 Median value: 100 % Median value: 61 % 250
200
150
100 The ADAMTS13 antigen and the activity values, show significant differences between normal and 50
TTP patients. % in Antigen ADAMTS-13 0 E019E.006 06/2019 TECHNOZYM TECHNOZYM TECHNOZYM Product ELISA TECHNOZYM Product [email protected] [email protected] +43 1 Phone: 86 373 67 Str. Brunner Arzneimitteln und Diagnostika GmbH Herstellung Technoclone von
® ® ® ®
ADAMTS13 Activity/Antigen ADAMTS13 ADAMTS13 INH ELISAADAMTS13 INH AntigenADAMTS13 ELISA Activity ELISA ADAMTS13 available on the TECHNOCLONE Youtube Channel (QR Channel Youtube TECHNOCLONE the on available video for Demonstration I 1230 1230 Vienna - 0 I
Fax: 1+43 86373 I www.technoclone.com I
Austria
TECHNOZYM
For information regarding our new ADAMTS13 activity screening assay screening activity ADAMTS13 new our regarding information For
1 x 12 ml TMB Substrate TMB 121 x ml Buffer Incubation 901 x ml Concentrate Buffer 801 x ml Wash anti 0.31 x ml strips ADAMTS13 12 8 x rec. Controls and Calibrators 0.57 x ml Solution Stop 121 x ml Substrate TMB 121 x ml Buffer Incubation 901 x ml Concentrate Buffer 801 x ml Wash anti 0.31 x ml strips ADAMTS13 anti 12 8 x Solution Stop 121 x ml GST 6 2 x ml Substrate TMB 121 x ml Concentrate Buffer 531 x ml Wash Buffer Reaction 301 x ml Conjugate HRP 121 x ml strips GST anti 12 8 x Contents 7 x 0.5 ml Calibrators and Controls and Calibrators 0.57 x ml Solution Stop 121 x ml 7 x 0.5 ml Calibrators and Controls and Calibrators 0.57 x ml Solution Stop 6 1 x ml Solution Peroxidase Stable 0.71 x ml Substrate Antigen 6 1 x ml Substrate Activity 2/1 3 x ml Buffer Incubation 901 x ml Concentrate Buffer 801 x ml Wash anti 0.31 x ml 8 anti 12/6 x Contents - 44 TECHNOSCREEN
®
ADAMTS13 Activity ELISA Activity ADAMTS13
-
ADAMTS13 strips ADAMTS13 - vWF73 Substrate vWF73 - - - IgG Conjugate Conjugate ADAMTS13 ADAMTS13 Conjugate ADAMTS13
® ADAMTS
ADAMTS13 Activity ADAMTS13 - Code)
ADAMTS13 Fluorogenic ELISAADAMTS13 Fluorogenic
-
13 Chromogenic ELISAs13
5450451 5450401 5450601 5450701 5450551 5450501 REF REF see separate folder separate see
48 tests 96 tests 96 tests 96 tests Tests 48 tests 482 x tests Tests