Dataset Information

Mass Spectrometric Analysis of Antibody-Epitope Peptide Complex Dissociation: Theoretical Concept and Practical Procedure of Binding Strength Characterization

ABSTRACT: We describe theoretical concepts and explain practical procedures of an electrospray mass spectrometry method, termed “Intact Transmission Epitope Mapping – Thermodynamic Weak-force Order (ITEM-TWO)”, by which can be determined apparent binding energies and dissociation constants of immune complex dissociation reactions in the gas phase. The protocol was developed using natural protein-ligand complexes (myoglobin and RNAse S) and was tested with two immune complexes (FLAG peptide - antiFLAG antibody and Troponin I epitope peptide - anti-Troponin I antibody). ITEM-TWO is a rapid method which requires very little sample consumption for identification of protein bound ligands and for determination of the gas phase protein-ligand complex binding strength.

INSTRUMENT(S): Synapt MS

ORGANISM(S): Equus Caballus (horse) Homo Sapiens (human)

SUBMITTER: Michael Kreutzer

LAB HEAD: Prof. Dr. Michael O. Glocker

PROVIDER: PXD021296 | Pride | 2020-10-29

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	04122017_MYOGLOBIN_NATIVE_DIL.raw.zip	Raw
	04122017_MYOGLOBIN_NATIVE_DIL_1.raw.zip	Raw
	04122017_MYOGLOBIN_NATIVE_DIL_2.raw.zip	Raw
	081018_RNASE_S_2.raw.zip	Raw
	091018_RNASE_S_2.raw.zip	Raw

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Publications

Mass Spectrometric Analysis of Antibody-Epitope Peptide Complex Dissociation: Theoretical Concept and Practical Procedure of Binding Strength Characterization.

Danquah Bright D BD Opuni Kwabena F M KFM Roewer Claudia C Koy Cornelia C Glocker Michael O MO

Molecules (Basel, Switzerland) 20201017 20

Electrospray mass spectrometry is applied to determine apparent binding energies and quasi equilibrium dissociation constants of immune complex dissociation reactions in the gas phase. Myoglobin, a natural protein-ligand complex, has been used to develop the procedure which starts from determining mean charge states and normalized and averaged ion intensities. The apparent dissociation constant KD m0g#= 3.60 × 10<sup>-12</sup> for the gas phase heme dissociation process was calculated from ...[more]

PMID: 33080923

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