PXD050732-2

PXD050732 is an original dataset announced via ProteomeXchange.

Title	Comparative analysis of laboratory-scale IgG purification methods from human serum
Description	Immunoglobulin G (IgG) purification is a critical process for evaluating its role in autoimmune diseases, which are defined by the occurrence of autoantibodies. Affinity chromatography with protein G is widely considered the optimal technique for laboratory-scale purification. However, this technique has some limitations, including the exposure of IgG to low pH, which can compromise the quality of the purified IgG. Here, we show that alternative methods for IgG purification are possible, while maintaining the quality of IgG. Different techniques for IgG purification from serum were evaluated and compared with protein G-based approaches: MelonTM Gel, caprylic acid-ammonium sulfate (CA-AS) precipitation, anion exchange chromatography with diethylaminoethyl (DEAE) following AS precipitation, and AS precipitation alone. The results demonstrated that the purification yield of these techniques surpassed that of protein G. However, differences in the purity of IgG were observed using GeLC-MS/MS. The avidity of purified IgG against selected targets (SARS-CoV-2 and topoisomerase-I) was similar between purified IgG obtained using all techniques and unpurified sera. Our work provides valuable insights for future studies of IgG function by recommending alternative purification methods that offer advantages in terms of yield, time efficiency, cost-effectiveness, and milder pH conditions than protein G.
HostingRepository	PRIDE
AnnounceDate	2024-10-22
AnnouncementXML	Submission_2024-10-22_06:57:39.715.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	BRAY FABRICE
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	monohydroxylated residue; deamidated residue; iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2024-03-18 11:17:07	ID requested
1	2024-09-02 09:03:53	announced
⏵ 2	2024-10-22 06:57:40	announced	2024-10-22: Updated project metadata.

10.1021/ACS.JPROTEOME.4C00268;

submitter keyword: Human Serum, Proteomics,Immunoglobulins G

fabrice bray
contact affiliation	​​Laboratoire MSAP UAR 3290 CNRS Bâtiment C4 Avenue Paul Langevin Cité Scientifique 59655 Villeneuve D'ascq 03 20 33 71 12 https://msap-lab.fr/
contact email	fabrice.bray@univ-lille.fr
lab head
BRAY FABRICE
contact affiliation	MSAP USR 3290
contact email	fabrice.bray@univ-lille.fr
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD050732
     1. Label: PRIDE project
     2. Name: Comparative analysis of laboratory-scale IgG purification methods from human serum