A new two-color Fab labeling method for autoantigen protein microarrays.
Antigen microarrays hold great promise for profiling the humoral immune response in the settings of autoimmunity, allergy and cancer. This approach involves immobilizing antigens on a slide surface and then exposing the array to biological fluids containing immunoglobulins. Although these arrays have proven extremely useful as research tools, they suffer from several sources of variability. To address these issues, we have developed a new two-color Fab labeling method that allows two samples to be applied simultaneously to the same array. This straightforward labeling approach improves reproducibility and reliably detects changes in autoantibody concentrations. Using this technique we profiled serum from a mouse model of systemic lupus erythematosus (SLE) and detected both expected and previously unrecognized reactivities. The improved labeling and detection method described here overcomes several problems that have hindered antigen microarrays and should facilitate translation to the clinical setting.
Kattah
Michael G
MG
Department of Medicine, Division of Immunology and Rheumatology, Stanford University School of Medicine, Stanford, California 94305, USA.
Alemi
Golnaz R
GR
Thibault
Donna L
DL
Balboni
Imelda
I
Utz
Paul J
PJ
D000818:Animals; D001323:Autoantibodies / Q000097:blood* / Q000737:chemistry / Q000276:immunology; D001324:Autoantigens / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D007140:Immunoglobulin Fab Fragments; D008180:Lupus Erythematosus, Systemic / Q000276:immunology*; D051379:Mice; D040081:Protein Array Analysis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D001323:Autoantibodies; D001324:Autoantigens; D007140:Immunoglobulin Fab Fragments