Kurzfassung

Western Blotting can effectively detect a specific antibody reactivity in a given sample. However, this analysis is time consuming, there is a lot of blot-to-blot variation, and is limited in sensitivity when used for mass screening of many samples simultaneously.
In comparison, a bead-based assay can be more usefull to capture immunoglobulin’s for downstream immunoprecipitation of wide range of proteins in a short period of time. The method will also be much much more sensitve since it is...Western Blotting can effectively detect a specific antibody reactivity in a given sample. However, this analysis is time consuming, there is a lot of blot-to-blot variation, and is limited in sensitivity when used for mass screening of many samples simultaneously.
In comparison, a bead-based assay can be more usefull to capture immunoglobulin’s for downstream immunoprecipitation of wide range of proteins in a short period of time. The method will also be much much more sensitve since it is based on mass spectrometry (MS).
Magnetic Protein G beads will be used for covalent cuppling of IgG from patient samples (e.g. serum samples). This covalent IgG-bead complexes can no be incubated with the antigens of interest. Then the bound antigens can be eluted through a pH-shift and the elutions will be measured on an MS. The antigen-incubation step will be repated several times. » weiterlesen» einklappen