Biotinylated platelets have an impaired response to agonists as evidenced by in vitro platelet aggregation tests.
Biotinylation of platelets, using a water soluble biotin analogue which reacts with primary amines, has been proposed to be a reliable technique for study of in vivo survival of platelets and their subpopulations. The information about the influence of this technique on platelet function has been limited. In the present work we studied the effect of in vitro biotinylation on platelet function and activation. Washed human platelets, at a concentration of 1 x 10(9)/L, were biotinylated with five different concentrations of sulfo-NHS-biotin or NHS-LC-biotin, ranging from 0 to 5 mM. The degree of platelet activation during and after biotinylation was monitored by measuring the externalization of P-selectin, and the platelet function was evaluated by aggregometry. It was observed that biotinylated platelets, in a dose dependent manner, displayed an impaired aggregation response. A slight increase in platelet membrane P-selectin occurred during the labelling procedure.
Magnusson
S
S
Department of Surgery, Sahlgrenska University Hospital, Göteborg, Sweden. stefan.magnusson@ss.gu.se
Hou
M
M
Hallberg
E C
EC
Breimer
M E
ME
Wadenvik
H
H
D000244:Adenosine Diphosphate / Q000494:pharmacology; D000322:Adrenergic Agonists / Q000494:pharmacology; D001710:Biotin / Q000031:analogs & derivatives* / Q000494:pharmacology; D019921:Biotinylation; D001792:Blood Platelets / Q000502:physiology*; D015153:Blotting, Western; D004305:Dose-Response Relationship, Drug; D004837:Epinephrine / Q000494:pharmacology; D005434:Flow Cytometry; D006490:Hemostatics / Q000494:pharmacology; D006801:Humans; D019007:P-Selectin / Q000096:biosynthesis / Q000187:drug effects; D010974:Platelet Aggregation / Q000187:drug effects*; D013268:Stimulation, Chemical; D013388:Succinimides / Q000494:pharmacology*; D013917:Thrombin / Q000494:pharmacology
D000322:Adrenergic Agonists; D006490:Hemostatics; D019007:P-Selectin; D013388:Succinimides; C065130:sulfo-N-hydroxysuccinimide-biotin; C061443:sulfosuccinimidyl 6-(biotinamido)hexanoate; D000244:Adenosine Diphosphate; D001710:Biotin; D013917:Thrombin; D004837:Epinephrine