Use of photografting technique for the immobilization of blood-grouping antibodies onto microtiter plates.
Commercial anti-A, anti-B, anti-A,B and anti-D monoclonal and polyclonal antisera were immobilized onto polystyrene microtiter plates using a photografting technique, to set up a new solid-phase assay (SPA) to be used for blood grouping. The reactivity and specificity of each grafted antisera were studied using red blood cells (RBCs) expressing normal and weak antigens. The stability of immobilized antisera was also studied. After dry storage of plates at +4 degrees C, room temperature, and +37 degrees C, SPA was performed using fresh and/or frozen RBCs. The same test was carried out after storing plates under protective conditions. Concordance of collected with expected results was obtained in all cases when the SPA was performed using monoclonal antisera and RBCs, with normal or weak expression of ABO and D antigens immediately after plate preparation or after dry storage at +4 degrees C. Plates stored dry at room temperature or at +37 degrees C gave inconsistent results, whereas a slight increase in reactivity was observed after storage under protective conditions. The specificity and the reactivity of tested antibodies were not modified by the immobilization procedure, not even after dry storage at +4 degrees C. Damage produced by water evaporation during dry storage in hard conditions could be reduced by adding a protective solution to microtiter wells at the beginning of storage.
D'Ambrosio
A
A
Centro Trasfusionale e di Immunologia dei Trapianti, IRCCS-Ospedale Maggiore Policlinico, Milano, Italy.
Rebulla
P
P
Revelli
N
N
Morelati
F
F
Marangoni
F
F
Sirchia
G
G
Bellobono
I R
IR
D000017:ABO Blood-Group System / Q000276:immunology*; D000911:Antibodies, Monoclonal; D001672:Biocompatible Materials; D001709:Biotechnology; D001788:Blood Grouping and Crossmatching / Q000379:methods*; D006801:Humans; D007158:Immunologic Techniques; D066298:In Vitro Techniques; D007518:Isoantibodies; D008422:Materials Testing; D010777:Photochemistry; D017319:Photosensitizing Agents
D000017:ABO Blood-Group System; D000911:Antibodies, Monoclonal; D001672:Biocompatible Materials; D007518:Isoantibodies; D017319:Photosensitizing Agents