Improvement of 2D-PAGE resolution of human, porcine and canine follicular fluid: comparison of two immunodepletion columns

Somayyeh Fahiminiya (1) , S. Roche (2, 3) , Nadine Gérard (1)

1 PRC - Physiologie de la reproduction et des comportements [Nouzilly]
2 IGH - Institut de génétique humaine
3 PPM - Pôle Protéome de Montpellier

Contents Follicular fluid provides the microenvironment within which somatic cells proliferate and differentiate, and the oocyte matures. It contains a number of soluble factors implicated in various stages of follicular development, most of them being functionally unknown. The presence of several high-abundance proteins, mainly originating from the blood circulation, is a major challenge of follicular fluid proteomic analysis, as these proteins can mask or decrease the visualization of follicle-specific proteins. In this study, we evaluated the efficiency of two immunodepletion columns (ProteomeLab (TM) IgY-HSA and MARS-6) on follicular fluids of human, porcine and canine prior to 2D-PAGE. Our results showed that both columns were suitable to remove some of the high-abundance proteins present in human and canine follicular fluid. In conclusion, we demonstrated that the immunodepletion strategy enables the detection of new protein spots, increases resolution and highly improves the intensity of low-abundance proteins by 2D-PAGE.

Mots clés: Proteomics; Follicular fluid; immunodepletion; 2D-PAGE; Canine; Human; Biomarker

Domaines: Autre [q-bio.OT]; Génomique, Transcriptomique et Protéomique [q-bio.GN]; Médecine vétérinaire et santé animale; Reproduction sexuée; Biochimie [q-bio.BM]

Somayyeh Fahiminiya, S. Roche, Nadine Gérard. Improvement of 2D-PAGE resolution of human, porcine and canine follicular fluid: comparison of two immunodepletion columns. Reproduction in Domestic Animals, 2012, 47 (5), pp.e67-e70. ⟨10.1111/j.1439-0531.2011.01939.x⟩. ⟨hal-01129661⟩