Self-assembled monolayers for MALDI-TOF mass spectrometry for immunoassays of human protein antigens.
This paper reports a method that combines self-assembled monolayers with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry to perform immunoassays on clinical samples. The immunosensors are prepared by immobilizing His-tagged protein G (or A) to a monolayer presenting the Ni2+ chelates, followed by immobilization of IgG antibodies with specificity for the intended analyte. The SAMDI mass spectrometry technique confirms the presence of the two proteins on the immunosensor and additionally provides a label-free analysis of antigens that bind to the sensor. This paper reports examples of detecting several proteins from human serum, including multianalyte assays that resolve each analyte according to their mass-to-charge ratio in the SAMDI spectra. An example is described wherein SAMDI is used to identify a proteolytic fragment of cystatin C in cerebral spinal fluids from patients diagnosed with multiple sclerosis. The SAMDI-TOF immunoassay, which combines well-defined surface chemistries for the selective and reproducible localization of analytes with mass spectrometry for label-free detection of analytes, may offer an alternative methodology to address many of the issues associated with standardized clinical diagnostics.
Patrie
Steven M
SM
Department of Pathology, University of Chicago, Chicago, Illinois 60637, USA.
Mrksich
Milan
M
D000941:Antigens / Q000097:blood* / Q000276:immunology; D002614:Chelating Agents / Q000737:chemistry; D002624:Chemistry, Clinical / Q000379:methods*; D055316:Cystatin C; D015891:Cystatins / Q000134:cerebrospinal fluid; D004800:Enzymes, Immobilized / Q000737:chemistry; D006639:Histidine / Q000737:chemistry; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007074:Immunoglobulin G / Q000276:immunology; D009103:Multiple Sclerosis / Q000134:cerebrospinal fluid; D009419:Nerve Tissue Proteins / Q000737:chemistry; D009532:Nickel / Q000737:chemistry; D011506:Proteins / Q000032:analysis* / Q000276:immunology / Q000378:metabolism*; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013205:Staphylococcal Protein A / Q000737:chemistry
D000941:Antigens; C525030:CST3 protein, human; D002614:Chelating Agents; D055316:Cystatin C; D015891:Cystatins; D004800:Enzymes, Immobilized; C029152:G-substrate; D007074:Immunoglobulin G; D009419:Nerve Tissue Proteins; D011506:Proteins; D013205:Staphylococcal Protein A; D006639:Histidine; D009532:Nickel