Linear epitope mapping by native mass spectrometry

Lu, Xiaojun; DeFelippis, Michael R.; Huang, Lihua

doi:10.1016/j.ab.2009.08.018

Cited by 27 publications

(22 citation statements)

References 23 publications

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“…Facing a collection of antibody candidates for one antigen, we may be able to categorize antibody binding at different epitopes with high throughput and low sample consumption. Although this approach does not locate the epitope, it may in the future by top-down sequencing [33, 34]. …”

Section: Resultsmentioning

confidence: 99%

Native Mass Spectrometry, Ion mobility, and Collision-Induced Unfolding Categorize Malaria Antigen/Antibody Binding

Huang

Salinas

Chen

et al. 2017

J. Am. Soc. Mass Spectrom.

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Plasmodium vivax Duffy Binding Protein (PvDBP) is a promising vaccine candidate for P. vivax malaria. Recently, we reported the epitopes on PvDBP region II (PvDBP-II) for three inhibitory monoclonal antibodies (2D10, 2H2, and 2C6). In this communication, we describe the combination of native mass spectrometry and ion mobility (IM) with collision induced unfolding (CIU) to study the conformation and stabilities of three malarial antigen-antibody complexes. These complexes, when collisionally activated, undergo conformational changes that depend on the location of the epitope. CIU patterns for PvDBP-II in complex with antibody 2D10 and 2H2 are highly similar, indicating comparable binding topology and stability. A different CIU fingerprint is observed for PvDBP-II/2C6, indicating that 2C6 binds to PvDBP-II on an epitope different from 2D10 and 2H2. This work supports the use of CIU as a means of classifying antigen-antibody complexes by their epitope maps in a high throughput screening workflow.

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Section: Resultsmentioning

confidence: 99%

Native Mass Spectrometry, Ion mobility, and Collision-Induced Unfolding Categorize Malaria Antigen/Antibody Binding

Huang

Salinas

Chen

et al. 2017

J. Am. Soc. Mass Spectrom.

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“…By contrast, other studies (Macht et al, ; Lyubarskaya et al, ; Kiselar & Downard, ; Lu, DeFelippis, & Huang, ; El‐Kased et al, ) describe in‐solution procedures, in which immobilization of neither antibody nor antigen prior to epitope mapping is necessary. These approaches provide non‐denaturing conditions throughout all experimental procedures and avoid the difficulties associated with lack of control of chemical side reactions, for example, during immobilization of the antibody (Al‐Majdoub et al, ).…”

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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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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“…[47]. Кроме того, определить В-эпитопы можно путём масс-спектрометрического анализа комплексов антиген-антитело, выявляя участок молекулы антигена, защищенный от протеолиза или модификации паратопом антитела [43,48]. Фрагменты молекул антигенов, обладающих Т-эпитопной активностью, выявляют по их способности вызывать пролиферацию Т-лимфоцитов в культуре [49].…”

Section: этапы создания синтетической пептидной вакциныunclassified

Synthetic peptide vaccines

Moisa

Колесанова

2011

Biomed Khim

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Linear epitope mapping by native mass spectrometry

Cited by 27 publications

References 23 publications

Native Mass Spectrometry, Ion mobility, and Collision-Induced Unfolding Categorize Malaria Antigen/Antibody Binding

Native Mass Spectrometry, Ion mobility, and Collision-Induced Unfolding Categorize Malaria Antigen/Antibody Binding

Mass spectrometric epitope mapping

Synthetic peptide vaccines

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