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001-es BibID:	BIBFORM054365
Első szerző:	Kappelmayer János (laboratóriumi szakorvos)
Cím:	Flow cytometric detection of intracellular myeloperoxidase, CD3 and CD79a : interaction between monoclonal antibody clones, fluorochromes and sample preparation protocols / Janos Kappelmayer, Jan W. Gratama, Eva Karaszi, Pablo Menendez, Juana Ciudad, Rosana Rivas, Alberto Orfao
Dátum:	2000
ISSN:	0022-1759
Megjegyzések:	Detection of intracellular myeloperoxidase (MPO), CD79a and CD3 has become the most specific tool for the assignmentof myeloid, B- and T-lymphoid lineages in acute leukemias. In order to establish the best combination of monoclonalantibody reagent and sample preparation technique for the intracellular detection of these three markers, we compared sixdifferent cell fixation?permeabilization kits (Cytofix/CytopermE, Fix and PermE, IntraprepE, IntrastainE, PermeacyteEand PermeafixE) using 12 fluorochrome conjugates derived from seven monoclonal antibody (mAb) clones. A total of 21samples corresponding to normal peripheral blood (n54), normal bone marrow (n53), acute myeloblastic leukemia (AML,n56), precursor B-acute lymphoblastic leukemia (ALL, n56) and T-ALL (n52) cases, were analysed in two centers. Allfixation / permeabilization methods resulted in decreased side scatter and mostly increased forward scatter as compared toerythrocyte-lyse-washed and 1% paraformaldehyde fixed samples. The autofluorescence levels of the leukocyte populationswas only significantly increased with use of the Cytofix/CytopermE kit and mildly with the other techniques. In addition,non-specific staining increased significantly for combinations of any anti-MPO mAb with the Cytofix/CytopermE kit andfor the CD3 clone S4.1 combined with any intracellular method. Anti-MPO antibodies gave a stronger fluorescence signalwhen conjugated to PE than when coupled to FITC. In conclusion, MPO-7-PE, UCHT-1-PE (CD3) and any HM57-PEconjugate (CD79a) in combination with Fix and PermE, IntraprepE, IntrastainE or PermeafixE, provided specific stainingof the respective markers in sufficient intensities. Thus, combined selection of fixation / permeabilization kits and monoclonalantibody reagents against CD3, CD79a and MPO is required for obtaining optimal cytoplasmic detection of these antigens.Ó 2000 Elsevier Science B.V. All rights reserved.
Tárgyszavak:	Orvostudományok Klinikai orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
Megjelenés:	Journal of Immunological Methods. - 242 : 1-2 (2000), p. 53-65. -
További szerzők:	Gratama, Jan Willem Karászi Éva Menendez, Pablo Ciudad, Juana Rivas, Rosana Orfao, Alberto
Internet cím:	Intézményi repozitóriumban (DEA) tárolt változat
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001-es BibID:	BIBFORM088736
035-os BibID:	(cikkazonosító)112877
Első szerző:	Szabó Gábor (PhD-hallgató)
Cím:	Distinct and overlapping effects of [beta]2-glycoprotein I conformational variants in ligand interactions and functional assays / Szabó Gábor, Pénzes Krisztina, Torner Bernadett, Fagyas Miklós, Tarr Tünde, Soltész Pál, Kis Gréta, Antal Miklós, Kappelmayer János
Dátum:	2020
ISSN:	0022-1759
Megjegyzések:	One of the most abundant coagulation proteins is ?2-glycoprotein I (?2GPI) that is present in humans at a concentration of around 200 mg/L. Its physiological role is only partially understood, but it adopts several different structural forms the majority of which are the open and closed forms. We isolated native (circular) ?2GPI and converted it into an open conformation. The effectiveness of these procedures was assessed by Western blot and negative-staining electron microscopy. We found that in coagulation assays the open form of ?2GPI had a significant prolonging effect on fibrin formation in a dilute prothrombin time test (p < 0.001). In the dilute activated partial thromboplastin time test, both conformations had a significant prolonging effect (p < 0.001) but the open conformation was more effective. In a fluorescent thrombin generation assay both conformations slightly delayed thrombin generation with no significant effect on the quantity of formed thrombin. By using surface plasmon resonance assays, the equilibrium dissociation constants of both the open and closed conformations of ?2GPI showed a similar and strong affinity to isolated anti-?2GPI autoantibodies (Kd closed ?2GPI = 5.17 ? 10-8 M, Kd open ?2GPI = 5.56 ? 10-8 M) and the open form had one order of magnitude stronger affinity to heparin (Kd = 0.30 ? 10-6 M) compared to the closed conformation (Kd = 3.50 ? 10-6 M). The two different forms of ?2GPI have distinct effects in functional tests and in ligand binding, which may considerably affect the intravascular events related to this abundant plasma protein in health and disease.
Tárgyszavak:	Orvostudományok Klinikai orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban folyóiratcikk Anti-[beta](2)-glycoprotein I Heparin Surface plasmon resonance Thrombin generation [beta](2)-glycoprotein I conformations
Megjelenés:	Journal of Immunological Methods. - 487 (2020), p. 112877. -
További szerzők:	Pénzes-Daku Krisztina (1978-) (biológus) Torner Bernadett (1997-) (molekuláris biológus) Fagyas Miklós (1984-) (orvos) Tarr Tünde (1976-) (belgyógyász, allergológus és klinikai immunológus) Soltész Pál (1961-) (belgyógyász, kardiológus) Kis Gréta (1979-) (molekuláris biológus) Antal Miklós (1951-) (orvos, anatómus) Kappelmayer János (1960-) (laboratóriumi szakorvos)
Pályázati támogatás:	OTKA K16 120725 OTKA GINOP-2.3.2-15-2016-00043 GINOP
Internet cím:	Szerző által megadott URL DOI Intézményi repozitóriumban (DEA) tárolt változat
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