Comparison of different chelates for lanthanide labeling of antibodies and application in a Western blot immunoassay combined with detection by laser ablation (LA-)ICP-MS

Waentig, Larissa; Jakubowski, Norbert; Hardt, Simone; Scheler, Christian; Roos, Peter H.; Linscheid, Michael

doi:10.1039/c2ja30068k

Cited by 57 publications

(77 citation statements)

References 35 publications

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“…Moreover, detection of five differentially labeled antibodies in one single Western blot assay is achieved (Waentig et al, ). Three commercially available labeling reagents (i.e., MeCAT, MAXPAR™, and SCN‐DOTA) were compared (Waentig et al, ). The results shows lower fmol range LOD can be reached in the Western blot immunoassay using MeCAT and MAXPAR™ as element labeling reagents, whereas even sub fmol LOD can be achieved in a dot blot experiment for the pure antibodies.…”

Section: La‐icpms‐based Immunoassay For Surface Analysismentioning

confidence: 99%

Inductively coupled plasma mass spectrometry‐based immunoassay: A review

Liu

Yang

et al. 2013

Mass Spectrometry Reviews

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The last 10 years witnessed the emerging and growing up of inductively coupled plasma mass spectrometry (ICPMS)-based immunoassay. Its high sensitivity and multiplex potential have made ICPMS a revolutionary technique for bioanalyte quantification after element-tagged immunoassay. This review focuses on the major developments and the applications of ICPMS-based immunoassay, with emphasis on methodological innovations. The ICPMS-based immunoassay with elemental tags of metal ions, nanoparticles, and metal containing polymers was discussed in detail. The recent development of multiplex assay, mass cytometry, suspension array, and surface analysis demonstrated the versatility and great potential of this technique. ICPMS-based immunoassay has become one of the key methods in bioanalysis.

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Section: La‐icpms‐based Immunoassay For Surface Analysismentioning

confidence: 99%

Inductively coupled plasma mass spectrometry‐based immunoassay: A review

Liu

Yang

et al. 2013

Mass Spectrometry Reviews

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“…Labeling via a maleimide group binds to the sulfhydryl residues after partial reduction of the antibody's cysteine‐based disulfide bridges. MeCAT and SCN‐DOTA contain a DOTA macro‐cycle (pink circle) to complex lanthanide ions whereas the MAXPAR™ polymer chain contains 4n DTPA residues as chelate (yellow circle)…”

Section: Applicationsmentioning

confidence: 99%

“…In cooperation with the group of Jakubowski, the partial reduction over a wide molar excess range was tested. The different resulting antibodies were analyzed in a Dotblot immunoassay and the best signal‐to‐noise (S/N) ratios and LODs were achieved with a 600‐fold molar excess . To quantify the antibody, which is pivotal for quantifying the antigen, either UV absorbance at 280 nm may be used (however, some labels could affect the measurement), or ICP‐MS and a Bradford assay is possible, all of them with limited accuracy .…”

Section: Applicationsmentioning

confidence: 99%

“…The different resulting antibodies were analyzed in a Dotblot immunoassay and the best signal-to-noise (S/N) ratios and LODs were achieved with a 600-fold molar excess. 66 To quantify the antibody, which is pivotal for quantifying the antigen, either UV absorbance at 280 nm may be used (however, some labels could affect the measurement), 65 or ICP-MS and a Bradford assay is possible, all of them with limited accuracy. 47,114 Most recently, we showed that using an intermediate Cu-free click system, antibodies can be labeled as well at the glycan structure without interference with the immune activity ( Figure 3I).…”

Section: Antibodiesmentioning

confidence: 99%

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Molecules and elements for quantitative bioanalysis: The allure of using electrospray, MALDI, and ICP mass spectrometry side‐by‐side

Linscheid

2018

Mass Spectrometry Reviews

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To understand biological processes, not only reliable identification, but quantification of constituents in biological processes play a pivotal role. This is especially true for the proteome: protein quantification must follow protein identification, since sometimes minute changes in abundance tell the real tale. To obtain quantitative data, many sophisticated strategies using electrospray and MALDI mass spectrometry (MS) have been developed in recent years. All of them have advantages and limitations. Several years ago, we started to work on strategies, which are principally capable to overcome some of these limits. The fundamental idea is to use elemental signals as a measure for quantities. We began by replacing the radioactive P with the "cold" natural P to quantify modified nucleotides and phosphorylated peptides and proteins and later used tagging strategies for quantification of proteins more generally. To do this, we introduced Inductively Coupled Plasma Mass Spectrometry (ICP-MS) into the bioanalytical workflows, allowing not only reliable and sensitive detection but also quantification based on isotope dilution absolute measurements using poly-isotopic elements. The detection capability of ICP-MS becomes particularly attractive with heavy metals. The covalently bound proteins tags developed in our group are based on the well-known DOTA chelate complex (1,4,7,10-tetraazacyclododecane-N,N',N″,N‴-tetraacetic acid) carrying ions of lanthanoides as metal core. In this review, I will outline the development of this mutual assistance between molecular and elemental mass spectrometry and discuss the scope and limitations particularly of peptide and protein quantification. The lanthanoide tags provide low detection limits, but offer multiplexing capabilities due to the number of very similar lanthanoides and their isotopes. With isotope dilution comes previously unknown accuracy. Separation techniques such as electrophoresis and HPLC were used and just slightly adapted workflows, already in use for quantification in bioanalysis. Imaging mass spectrometry (MSI) with MALDI and laser ablation ICP-MS complemented the range of application in recent years.

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“…383 A special focus was set to apply the multielement capabilities of ICP-MS for the design of multiparametric immunoassays. In particular, the lanthanide tagging of antibodies with metal-coded affinity tags (MeCAT) has been investigated and optimised for application in a Western blot analysed by LA-ICP-MS for which a particular laser-ablation cell was designed.…”

Section: Element-specific Detection Of Small Molecules In Biological mentioning

confidence: 99%

Inductively Coupled Plasma Mass Spectrometry

Jakubowski

Horsky

Roos

et al. 2014

Sector Field Mass Spectrometry for Elemental and Isotopic Analysis

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ICP-MS is based on the formation of (preferentially monovalent positively) charged atomic ions in an inductively coupled Ar plasma at almost 10 000 K. The ions formed are transferred from the plasma source at ambient pressure into a mass separator operated at high vacuum via a set of cones. The ions are separated according to their mass/charge ratio in the mass separator (quadrupole, magnetic sector field or time-of-flight mass separator). In most cases, the ions are detected using a secondary electron multiplier; in some set-ups (also) a Faraday cup can be used. Single-collector (scanning mass spectrometer usually used for quantitative elemental analysis) or multicollector (static operation of mass spectrometer for precise isotope ratio analysis) configurations can be found (Figures 12.1 and 12.2).

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Comparison of different chelates for lanthanide labeling of antibodies and application in a Western blot immunoassay combined with detection by laser ablation (LA-)ICP-MS

Cited by 57 publications

References 35 publications

Inductively coupled plasma mass spectrometry‐based immunoassay: A review

Inductively coupled plasma mass spectrometry‐based immunoassay: A review

Molecules and elements for quantitative bioanalysis: The allure of using electrospray, MALDI, and ICP mass spectrometry side‐by‐side

Inductively Coupled Plasma Mass Spectrometry

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