reversible platelet stabilization - agro-bio · reversible platelet stabilization ... aggregation...
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Reversible platelet stabilizationUsing Apyrase ADP-High and Apyrase ADP-Premium for:
ADP and ATP degradation…
Standardization of platelet washing protocols
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5
2 allée de la Chavannerie – 45240 La Ferté Saint-Aubin – Francewww.agro-bio.com
Contact us at +33 2 38 64 83 50 or on [email protected]
Product references Apyrase ADP-High Apyrase ADP-Premium
Cat. # PY0623100 PY0623200 PY0623500 PY0627100 PY0627200 PY0627500
Packaging 1 fl. x 100 U 1 fl. x 200 U 1 fl. x 500 U 1 fl. x 100 U 1 fl. x 200 U 1 fl. x 500 U
Lyophilized products.1,000 and 2,000 U packaging are solely available frozen and shipped on dry-ice (upon request only).
Washed platelet aggregation study induced by ADP
From JP Cazenave et al., 2004, Preparation of washed platelet suspension from human and rodent blood.
From E. Maurer’s thesis, 2014, Role of initial steps of platelet adhesion and importance of pulsatile flow in platelet aggregation.
Kindly provided by UMR_S949, EFS ALCA, France.
Apyrase ADP-HighApyrase ADP-Premium
Washed platelet stability versus platelet-rich plasma stability
Features Apyrase ADP-High Apyrase ADP-Premium
Specific ADP activity > 20 U/mg > 200 U/mgPurification By precipitation By affinity
ApplicationPrimary hemostasisApyrase offers a higher specific ADP activity than other commercial apyrases. Apyrase ADP is more suitable for platelet studies.
~ 3 hours for PRP 5 to 8 hours for isolated washed platelet
Platelet shape change
Results of platelet aggregation
Resting state, discoid shape Platelet aggregation Platelet disaggregation
Aggregation of washed platelets induced by ADP at 5 µM
Platelets resuspended in buffer initially without Apyrase ADP, then addition
of Apyrase ADP before ADP
Platelets resuspended in bufferwithout Apyrase ADP
1 2 3
Platelets resuspended in bufferwith Apyrase ADP
20%
0%
40%
90%
Light transmission [%]
Time
Fibr
inog
en
AD
P
1
2
3
Principle Adding Apyrase ADP to a washed platelet suspension maintains platelet reactivity by:
Preventing ADP P2Y1 and ATP P2X1 platelet receptor desensitization,
Maintaining platelet discoid shape.
Apyrase ADP degrades the nucleotides released during platelet washing:
Washed platelets
OHOH
OH N
N
NH2
N
NP OO
OO
+ Pi
AMP + PiADP + PiATP
OHOH
OH N
N
NH2
N
NOH
HO P P OOO
OO
Apyrase
OHOH
O N
N
H2N
N
NOP P O
OOOO
OPO
OO
Apyrase
+ Pi
P2X1 P2Y1 P2Y12
ATP ADP ADP
No Aggregation
Receptor desensitization
ATP
P2X1
ADP
P2Y1 P2Y12
Without apyrase
ADP
With apyrase
Aggregation
Full receptor sensitization
P2X1 P2Y1 P2Y12
ATP ADP ADP
No Aggregation
Receptor desensitization
ATP
P2X1
ADP
P2Y1 P2Y12
Without apyrase
ADP
With apyrase
Aggregation
Full receptor sensitization
20%
0%
40%
90%
Light transmission [%]
Time
Fibr
inog
en
AD
P
20%
0%
40%
90%
Light transmission [%]
Time
Fibr
inog
en
AD
P