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Distinct Binding Characteristics of Pathogenic Anti-Platelet Factor-4/Polyanion Antibodies to Antigens Coated on Different Substrates: A Perspective on Clinical Application.
ACS Nano 2018 December 13
The polyanion heparin, which is frequently used in patients, complexes with the platelet-derived cationic chemokine platelet factor (PF4, CXCL4). This results in the formation of anti-PF4/heparin antibodies (anti-PF4/H Abs). Anti-PF4/H Abs are classified into three groups: (i) nonpathogenic Abs (group 1) with no clinical relevance; (ii) pathogenic heparin-dependent Abs (group 2), which activate platelets and can cause the severe adverse drug effect heparin-induced thrombocytopenia (HIT); and (iii) pathogenic autoimmune-HIT Abs (group 3), in which group 3 anti-PF4/H Abs causes a HIT-like autoimmune disease in the absence of heparin. Enzyme immunoassays using PF4/H complexes coated on the solid phase for detection of anti-PF4/H Abs cannot differentiate between pathogenic and nonpathogenic anti-PF4/H Abs. By single-molecule force spectroscopy, we identify a specific feature of pathogenic group 2 and group 3 Abs antibodies that (in contrast to nonpathogenic group 1 Abs) their binding forces to PF4/H complexes coated on platelets were significantly higher compared with those of PF4/H complexes immobilized on a solid phase. Only group 3 Abs showed high binding forces to platelets without the addition of PF4. In the presence of 50 μg/mL PF4, group 2 Abs also showed high binding forces to platelets. In contrast, binding forces of group 1 Abs always remained low (<100 pN). Our findings may have major relevance for the development of clinically applicable solid-phase assays, which allow differentiation of pathogenic platelet-activating from nonpathogenic anti-PF4/H Abs. Membrane-based expression of antigens might also increase the specificity of other assays for the detection of pathogenic (auto)-antibodies in clinical medicine.
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