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Immunochemical characterization of platelet‐specific alloantigens
Author(s) -
Mulder Arend,
Leeuwen Eleonora F.,
Veenboer Geertruida J. M.,
Tetteroo Pedro A. T.,
Borne Albert E. G. Kr.
Publication year - 1984
Publication title -
scandinavian journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.904
H-Index - 84
eISSN - 1600-0609
pISSN - 0036-553X
DOI - 10.1111/j.1600-0609.1984.tb02227.x
Subject(s) - platelet , characterization (materials science) , computational biology , immunology , biology , materials science , nanotechnology
Immunoprecipitation was performed with platelet‐specific alloantisera (anti‐Zw a , ‐Zw b , ‐Bak a and antiserum Luc) and 125 I‐labelled platelets of a panel of donors typed for these platelet‐specific alloantigens. This was done by sensitization of intact, radiolabelled platelets with the antisera, solubilization of the sensitized platelets with Nonidet P‐40 and recovery of the immune complexes from the lysate with heat‐killed, formalin‐fixed Staphylococcus aureus, strain Cowan I. In the case of platelet antibodies of the IgM class, bacteria also preincubated with rabbit‐IgG anti‐human IgM before treatment of the lysate were applied. Radiolabelled antigens were then eluted from the bacteria by boiling in an SDS‐mercaptoethanol‐containing buffer. Subsequently, the eluted antigens were analyzed by SDS‐polyacrylamide gel electrophoresis. It revealed that both the Zw a and the Bak a antigens are located on the glycoprotein‐IIb/IIIa complex. The location of the Zw b antigen could not be established in this way. From the intensity of the precipitated glycoprotein bands, it appeared that Zw a is probably a marker of glycoprotein IIIa, Bak a or glycoprotein IIb. However, immuno‐precipitates of anti‐Zw a and anti‐Bak a sera both carried, to some extent, complexes of glycoproteins IIb and IIIa. Iodinated platelets of a Glanzmann thrombasthenia patient failed to show any material precipitated by these antisera or the antiserum Luc, known to contain antibodies against both glycoproteins.