Dataset Information

Online electrochemical reduction of both inter- and intramolecular disulfide bridges in immunoglobulins

ABSTRACT: Cysteine disulfide bridges can be reduced by chemical methods, like an incubation with DTT. However, reduction with an electrochemical method has also been demonstrated. Electrochemical reduction can be implemented online, after LC separation and before mass spectrometry. For the study of antibody molecules, reduction of the intermolecular disulfide bridges between heavy and light chains has been demonstrated in literature. However, the reduction of intramolecular disulfide bridges has remained elusive. Here, we demonstrate the online electrochemical reduction of a therapeutic monoclonal antibody. The complete reduction of these molecules will facilitate the study of recombinant or natural antibodies by MS and MS/MS.

INSTRUMENT(S): Orbitrap Fusion Lumos, Synapt MS

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): B Cell

SUBMITTER: Martijn van Duijn

LAB HEAD: Theo Marten Luider

PROVIDER: PXD021649 | Pride | 2022-05-27

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	190730OLc1_MvD-1939-GdnDTT_C012.raw	Raw
	190730OLc1_MvD-1939-Intact1000mV_C017.raw	Raw
	190730OLc1_MvD-1939-Intact600mV_C015.raw	Raw
	190730OLc1_MvD-1939-TrisDTT_C013.raw	Raw
	1939AvaLCPepXML.xml	Xml

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Publications

Online Electrochemical Reduction of Both Inter- and Intramolecular Disulfide Bridges in Immunoglobulins.

Vanduijn Martijn M MM Brouwer Hendrik-Jan HJ Sanz de la Torre Pablo P Chervet Jean-Pierre JP Luider Theo M TM

Analytical chemistry 20220204 7

Electrochemical reduction of intermolecular disulfide bridges has previously been demonstrated in immunoglobulins but failed to achieve reduction of intramolecular bonds. We now report an improved method that achieves the full reduction of both intermolecular and intramolecular disulfide bridges in a set of monoclonal antibodies based on their intact mass and on MS/MS analysis. The system uses an online electrochemical flow cell positioned online between a chromatography system and a mass spectr ...[more]

PMID: 35119270

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