Dataset Information

IP-to-MS: an unbiased workflow for antigen profiling

ABSTRACT: Immunoprecipitation is among the most widely utilized methods in biomedical research, with applications that include the identification of antibody targets and associated proteins. The path to identifying these targets is not straightforward, however, and often requires the use of chemical crosslinking and/or gel electrophoresis to separate targets from an overabundance of immunoglobulin protein. Such experiments are labor intensive and often yield long lists of candidate antibody targets. Here, we describe an unbiased immunoprecipitation-to-mass spectrometry (IP-to-MS) method that relies on a novel protein tag to separate low abundance immunoprecipitated proteins from overwhelmingly abundant immunoglobulins. We demonstrate that the IP-to-MS serotyping workflow is highly reproducible and can be used for the identification of novel, patient-specific antigen targets in multiple disease states. Furthermore, we show that IP-to-MS may outperform conventional methods of antibody detection, including ELISA, while also enabling patient stratification beyond what is possible with traditional approaches.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Blood Cell, Blood Serum

DISEASE(S): Interstitial Lung Disease,Connective Tissue Disease,Myositis,Systemic Scleroderma

SUBMITTER: Stephanie Biedka

LAB HEAD: Jonathan Minden

PROVIDER: PXD056617 | Pride | 2025-05-07

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	Gel1-26_Jan2024.rar	Other
	Gel1-26_Samples_Master_Jan2024.xlsx	Xlsx
	IP1-37_Oct2023.rar	Other
	IP1-37_ProteinList_Oct2023.xlsx	Xlsx
	Minden_051822_Results.xlsx	Xlsx

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Publications

IP-to-MS: An Unbiased Workflow for Antigen Profiling.

Biedka Stephanie S Yablonska Svitlana S Peng Xi X Alkam Duah D Hartoyo Mara M VanEvery Hannah H Kass Daniel J DJ Byrum Stephanie D SD Xiao Kunhong K Zhang Yingze Y Domsic Robyn T RT Lafyatis Robert R Ascherman Dana P DP Minden Jonathan S JS

Journal of proteome research 20250115 2

Immunoprecipitation is among the most widely utilized methods in biomedical research, with applications that include the identification of antibody targets and associated proteins. The path to identifying these targets is not straightforward, however, and often requires the use of chemical cross-linking and/or gel electrophoresis to separate targets from an overabundance of immunoglobulin protein. Such experiments are labor intensive and often yield long lists of candidate antibody targets. Here ...[more]

PMID: 39814365

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