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To study VITT, patient samples are crucial but have become a scarce resource. Recombinant antibodies (rAbs) derived from VITT patient characteristic amino acid sequences of anti-PF4 IgG are an alternative to study VITT pathophysiology. Methods: Amino acid sequences of the variable region of immunoglobulin light and heavy chain of anti-PF4 IgG derived from VITT patients were obtained by mass spectrometry sequencing and rAbs were synthetized by reverse-engineering. Six different rAbs were produced: CR23003, CR23004, and CR23005 (from a patient vaccinated with Jcovden, Johnson &amp; Johnson-Janssen (Beerse, Belgium)), CR22046, and CR22050 and CR22066 (from two different patients vaccinated with Vaxzevria, AstraZeneca (Cambridge, UK)). These rAbs were further characterized using anti-PF4 and anti-PF4\/heparin IgG ELISAs, rapid anti-PF4 and anti-PF4\/polyanion chemiluminescence assays, and PF4-induced platelet activation assay (PIPA) and their capacity to induce procoagulant platelets. Results: rAbs bound to PF4 alone, but not to PF4\/polyanion complexes in rapid chemiluminescence assays. Chemiluminescence assays and both anti-PF4 IgG and anti-PF4 IgG\/heparin ELISA showed concentration-dependent PF4 binding of all six rAbs, however, with different reactivities among them. PIPA showed a similar, concentration-dependent platelet activation pattern. rAbs varied in their reactivity and the majority of the tested rAbs were able to induce procoagulant platelets. Conclusions: The six rAbs derived from VITT patients reflect VITT-typical binding capacities and the ability to activate platelets. Therefore, these rAbs offer an attractive new option to study VITT pathophysiology.<\/jats:p>","DOI":"10.3390\/vaccines13010003","type":"journal-article","created":{"date-parts":[[2024,12,24]],"date-time":"2024-12-24T09:00:57Z","timestamp":1735030857000},"page":"3","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Recombinant Anti-PF4 Antibodies Derived from Patients with Vaccine-Induced Immune Thrombocytopenia and Thrombosis (VITT) Facilitate Research and Laboratory Diagnosis of VITT"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7942-3273","authenticated-orcid":false,"given":"Luisa","family":"M\u00fcller","sequence":"first","affiliation":[{"name":"Institut f\u00fcr Transfusionsmedizin, Universit\u00e4tsmedizin Greifswald, 17489 Greifswald, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8140-4335","authenticated-orcid":false,"given":"Venkata A. S.","family":"Dabbiru","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Transfusionsmedizin, Universit\u00e4tsmedizin Greifswald, 17489 Greifswald, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lucy","family":"Rutten","sequence":"additional","affiliation":[{"name":"Janssen Vaccines & Prevention BV, 2333 CN Leiden, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rinke","family":"Bos","sequence":"additional","affiliation":[{"name":"Janssen Vaccines & Prevention BV, 2333 CN Leiden, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2822-6231","authenticated-orcid":false,"given":"Roland","family":"Zahn","sequence":"additional","affiliation":[{"name":"Janssen Vaccines & Prevention BV, 2333 CN Leiden, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefan","family":"Handtke","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Transfusionsmedizin, Universit\u00e4tsmedizin Greifswald, 17489 Greifswald, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0177-6508","authenticated-orcid":false,"given":"Thomas","family":"Thiele","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Transfusionsmedizin, Universit\u00e4tsmedizin Greifswald, 17489 Greifswald, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marta","family":"Palicio","sequence":"additional","affiliation":[{"name":"Werfen, Lli\u00e7\u00e0 d\u2019Amunt, 08186 Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-1846-4526","authenticated-orcid":false,"given":"Olga","family":"Esteban","sequence":"additional","affiliation":[{"name":"Werfen, Lli\u00e7\u00e0 d\u2019Amunt, 08186 Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4524-4290","authenticated-orcid":false,"given":"Marta","family":"Broto","sequence":"additional","affiliation":[{"name":"Werfen, Lli\u00e7\u00e0 d\u2019Amunt, 08186 Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tom Paul","family":"Gordon","sequence":"additional","affiliation":[{"name":"Department of Immunology, College of Medicine and Public Health, Flinders University and SA Pathology, Bedford Park, Adelaide, SA 5042, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8343-7336","authenticated-orcid":false,"given":"Andreas","family":"Greinacher","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Transfusionsmedizin, Universit\u00e4tsmedizin Greifswald, 17489 Greifswald, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing Jing","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Immunology, College of Medicine and Public Health, Flinders University and SA Pathology, Bedford Park, Adelaide, SA 5042, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6660-9949","authenticated-orcid":false,"given":"Linda","family":"Sch\u00f6nborn","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Transfusionsmedizin, Universit\u00e4tsmedizin Greifswald, 17489 Greifswald, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,12,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"198114","DOI":"10.1016\/j.virusres.2020.198114","article-title":"COVID-19 Vaccine: A comprehensive status report","volume":"288","author":"Kaur","year":"2020","journal-title":"Virus Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2092","DOI":"10.1056\/NEJMoa2104840","article-title":"Thrombotic Thrombocytopenia after ChAdOx1 nCov-19 Vaccination","volume":"384","author":"Greinacher","year":"2021","journal-title":"N. 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