A simple and practical agglutination assay for the detection of human anti-mouse antibodies.
OBJECTIVE
Human anti-mouse antibodies (HAMAs) are relatively common in human serum and may interfere with therapeutic and diagnostic mouse monoclonal antibodies (MoAbs). We developed a simple particle agglutination test (PaGIA) for the detection of HAMAs.


METHODS
Red-dyed high density particles were coated with monoclonal mouse IgG. These particles were incubated in the reaction chamber of a gel-card together with serum samples obtained from healthy blood donors (n=32), and patients with clinically proven autoimmune thrombocytopenia (AITP; n=26). Positive reactions were defined by a layer of particles on top of the gel or agglutinated particles dispersed throughout the gel matrix. Furthermore, MoAb-coated particles were subjected to flow cytometry and the results were compared with the new HAMA PaGIA.


RESULTS
HAMAs were detectable in 33% of serum samples tested (n=58). Results from flow cytometric analysis revealed a high parallel to those obtained by the PaGIA. Interestingly, we observed an increased incidence of HAMAs in AITP patients (42%) compared to healthy blood donors (26%).


CONCLUSIONS
The new HAMA PaGIA allows a specific and easy, rapid detection of HAMAs and is suitable for large scale testing.
Socher
Ines
I
Institute for Clinical Immunology and Transfusion Medicine, Justus Liebig University, Giessen, Germany.
Meyer
Oliver
O
Aslan
Tunay
T
Bombard
Stéphane
S
Kiesewetter
Holger
H
Salama
Abdulgabar
A
D000372:Agglutination Tests / Q000379:methods*; D000818:Animals; D000910:Antibodies, Heterophile / Q000097:blood*; D000911:Antibodies, Monoclonal / Q000097:blood*; D001709:Biotechnology / Q000379:methods; D005434:Flow Cytometry; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood*; D051379:Mice / Q000276:immunology*; D016553:Purpura, Thrombocytopenic, Idiopathic / Q000175:diagnosis / Q000276:immunology; D012680:Sensitivity and Specificity
D000910:Antibodies, Heterophile; D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G