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Comparative analysis of laboratory-scale IgG purification methods from human serum

ABSTRACT: 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.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Blood Plasma

SUBMITTER: BRAY FABRICE

LAB HEAD: fabrice bray

PROVIDER: PXD050732 | Pride | 2024-09-02

REPOSITORIES: Pride

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			Action	DRS
	1.msf	Msf
	1.raw	Raw
	10.msf	Msf
	10.raw	Raw
	100.msf	Msf

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