Next generation of labeling reagents for quantitative and multiplexing immunoassays by use of LA-ICP-MS
(English)Manuscript (preprint) (Other academic)
Abstract. Immuno imaging by use of laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) is a growing research field in the life sciences as biology and biomedicine. Various element labeling strategies for antibodies have been developed for the application of multiplex immunoassays analyzed by use of LA-ICP-MS. High multiplexing capabilities, a wide linear dynamic range and the possibility of absolute quantification are the main advantages of ICP-MS. But in the context of immuno imaging by use of LA-ICP-MS quantification of analytes is limited due to a non-controllable antibody labeling chemistry. in the presented proof-of-principle A novel antibody labeling technique has been investigated which results in a controlled labeling degree. A small affinity protein based on the C2 domain of protein G was modified with conventional metal coded tags (MeCAT) after introducing a cysteine in the C-terminus of the protein. The modified C2 domain photo crosslinks to the Fc or Fab region of the IgG and allows specific and covalent labeling of antibodies for multiplex immunoassays analyzed by use of LA-ICP-MS. In combination with a house-made calibration membrane the amount of labeled antibody-antigen complexes in a multiplex Western Blot immunoassay was determined by LA-ICP-MS.
Engineering and Technology
IdentifiersURN: urn:nbn:se:kth:diva-191160OAI: oai:DiVA.org:kth-191160DiVA: diva2:955165