Mass Spectrometry

Przybylski, Michael

doi:10.1002/9783527654703.ch11

Cited by 2 publications

(7 citation statements)

References 172 publications

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“…The aim of these developments is to streamline in‐solution experimental steps in such a way that on‐line coupling to mass spectrometers becomes feasible. To this end an interface is being developed that connects a biosensor fluid system to an ESI source with an internal desalting device (Przybylski, ).…”

Section: Discussionmentioning

confidence: 99%

“…Because the success of mass spectrometric epitope mapping was more connected with liquid handling and sample preparation improvements, rather than with mass spectrometric ionization conditions or mass spectrometric analyzer technologies, the early mass spectrometers presented a limitation only with respect to sensitivity of detection. The tremendous advancements in mass spectrometry instrumentation, for example, concerning ion transmission and detection, have resulted in more sensitive, more accurate, and more reliable analyses of biomolecules, such as proteins and peptides (Parker, Warren, & Mocanu, ; Przybylski, ). Concomitantly, the development of the two soft ionization methods MALDI and ESI (Wilm, ; Touboul & Zenobi, ) have provided dramatic improvements in sensitivity for peptide and protein analysis.…”

Section: Epitope Excision and Epitope Extractionmentioning

confidence: 99%

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Mass spectrometric epitope mapping

Opuni

Al-Majdoub

Yefremova

et al. 2016

Mass Spectrometry Reviews

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Mass spectrometric epitope mapping has become a versatile method to precisely determine a soluble antigen's partial structure that directly interacts with an antibody in solution. Typical lengths of investigated antigens have increased up to several 100 amino acids while experimentally determined epitope peptides have decreased in length to on average 10-15 amino acids. Since the early 1990s more and more sophisticated methods have been developed and have forwarded a bouquet of suitable approaches for epitope mapping with immobilized, temporarily immobilized, and free-floating antibodies. While up to now monoclonal antibodies have been mostly used in epitope mapping experiments, the applicability of polyclonal antibodies has been proven. The antibody's resistance towards enzymatic proteolysis has been of key importance for the two mostly applied methods: epitope excision and epitope extraction. Sample consumption has dropped to low pmol amounts on both, the antigen and the antibody. While adequate in-solution sample handling has been most important for successful epitope mapping, mass spectrometric analysis has been found the most suitable read-out method from early on. The rapidity by which mass spectrometric epitope mapping nowadays is executed outperforms all alternative methods. Thus, it can be asserted that mass spectrometric epitope mapping has reached a state of maturity, which allows it to be used in any mass spectrometry laboratory. After 25 years of constant and steady improvements, its application to clinical samples, for example, for patient characterization and stratification, is anticipated in the near future. © 2016 Wiley Periodicals, Inc. Mass Spec Rev 37:229-241, 2018.

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Section: Discussionmentioning

confidence: 99%

Section: Epitope Excision and Epitope Extractionmentioning

confidence: 99%

Mass spectrometric epitope mapping

Opuni

Al-Majdoub

Yefremova

et al. 2016

Mass Spectrometry Reviews

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“…In recent years, antibodies have found wide-spread application for diagnostics as well as therapy, and currently constitute the fastest growing group of medication with sales volumes up to USD 100 billion (in 2018) [ 1 , 2 , 3 , 4 ]. Antibodies also contribute a dominant share to disease diagnostics, being almost indispensable in laboratory assays, such as ELISA, Western Blot, and biosensor analysis [ 4 , 5 , 6 ]. Biosensor techniques using antibodies, such as surface plasmon resonance (SPR) are well established for detection and quantification of biomolecular interactions.…”

Section: Introductionmentioning

confidence: 99%

“…Biosensor techniques using antibodies, such as surface plasmon resonance (SPR) are well established for detection and quantification of biomolecular interactions. Although mass spectrometry (MS) is an established tool for molecular characterization of biomolecular interactions [ 6 , 7 ], the combination of biosensor technologies and mass spectrometry is still in an initial state of development and application [ 6 , 7 , 8 , 9 ]. Recently, biosensor and mass spectrometry technologies are gaining increasing interest as “hybrid” tools for detection, chemical structure determination and quantification of protein–antibody interactions, particularly recognition epitopes of antibodies [ 4 , 6 , 7 , 9 ].…”

Section: Introductionmentioning

confidence: 99%

“…The molecular characterization of antibody epitopes of a peptide or protein antigen is a key element for the discovery of biomarkers, vaccines and in drug development [ 4 , 6 , 7 , 10 , 11 , 12 ]. It is well established that antibodies bind to different peptide or protein ligands via linear (sequence) or assembled (“conformational”) epitopes [ 4 ].…”

Section: Introductionmentioning

confidence: 99%

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Identification and Affinity Determination of Protein-Antibody and Protein-Aptamer Epitopes by Biosensor-Mass Spectrometry Combination

Lupu

Wiegand

Holdschick

et al. 2021

IJMS

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Analytical methods for molecular characterization of diagnostic or therapeutic targets have recently gained high interest. This review summarizes the combination of mass spectrometry and surface plasmon resonance (SPR) biosensor analysis for identification and affinity determination of protein interactions with antibodies and DNA-aptamers. The binding constant (KD) of a protein–antibody complex is first determined by immobilizing an antibody or DNA-aptamer on an SPR chip. A proteolytic peptide mixture is then applied to the chip, and following removal of unbound material by washing, the epitope(s) peptide(s) are eluted and identified by MALDI-MS. The SPR-MS combination was applied to a wide range of affinity pairs. Distinct epitope peptides were identified for the cardiac biomarker myoglobin (MG) both from monoclonal and polyclonal antibodies, and binding constants determined for equine and human MG provided molecular assessment of cross immunoreactivities. Mass spectrometric epitope identifications were obtained for linear, as well as for assembled (“conformational”) antibody epitopes, e.g., for the polypeptide chemokine Interleukin-8. Immobilization using protein G substantially improved surface fixation and antibody stabilities for epitope identification and affinity determination. Moreover, epitopes were successfully determined for polyclonal antibodies from biological material, such as from patient antisera upon enzyme replacement therapy of lysosomal diseases. The SPR-MS combination was also successfully applied to identify linear and assembled epitopes for DNA–aptamer interaction complexes of the tumor diagnostic protein C-Met. In summary, the SPR-MS combination has been established as a powerful molecular tool for identification of protein interaction epitopes.

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Mass Spectrometry

Cited by 2 publications

References 172 publications

Mass spectrometric epitope mapping

Mass spectrometric epitope mapping

Identification and Affinity Determination of Protein-Antibody and Protein-Aptamer Epitopes by Biosensor-Mass Spectrometry Combination

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