Next generation of labeling reagents for quantitative and multiplexing immunoassays by the use of LA-ICP-MS.
Immuno imaging by the use of Laser Ablation Inductively Coupled Mass Spectrometry (LA-ICP-MS) is a growing research field in life sciences such as biology and biomedicine. Various element labeling strategies for antibodies have been developed for the application of multiplex immunoassays analyzed by the 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 the use of LA-ICP-MS, quantification of analytes is limited due to 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 into 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 immunoassay analysis by the 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.
Alba Nova University Center, KTH-Royal Institute of Technology, Division of Protein Technology, Stockholm, Sweden.
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http://orcid.org/0000-0002-4751-2519
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Kanje
Bundesanstalt für Materialforschung und-prüfung (BAM), 1.1 Division Inorganic Trace Analysis, Richard-Willstätter-Str. 11, 12489 Berlin, Germany. larissa.mueller@thermofisher.com and Humboldt-Universität zu Berlin, Institut für Chemie, Brook-Taylor-Straße 2, 12489 Berlin, Germany.
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Herrmann
Alba Nova University Center, KTH-Royal Institute of Technology, Division of Protein Technology, Stockholm, Sweden.
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Hober
Bundesanstalt für Materialforschung und-prüfung (BAM), 1.1 Division Inorganic Trace Analysis, Richard-Willstätter-Str. 11, 12489 Berlin, Germany. larissa.mueller@thermofisher.com.
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Mueller
D000906:Antibodies / Q000737:chemistry*; D007118:Immunoassay; D007202:Indicators and Reagents / Q000737:chemistry*; D007834:Lasers; D013058:Mass Spectrometry; D008670:Metals; D000075082:Proof of Concept Study
D000906:Antibodies; D007202:Indicators and Reagents; D008670:Metals