Compatibility of Distinct Label-Free Proteomic Workflows in Absolute Quantification of Proteins Linked to the Oocyte Quality in Human Follicular Fluid

We present two separate label-free quantitative workflows based on different high-resolution mass spectrometers and LC setups, which are termed after the utilized instrument: Quad-Orbitrap (nano-LC) and Triple Quad-TOF (micro-LC) and their directed adaptation toward the analysis of human follicular fluid proteome. We identified about 1000 proteins in each distinct workflow using various sample preparation methods. With assistance of the Total Protein Approach, we were able to obtain absolute protein concentrations for each workflow. In a pilot study of twenty samples linked to diverse oocyte quality status from four donors, 455 and 215 proteins were quantified by the Quad-Orbitrap and Triple Quad-TOF workflows, respectively. The concentration values obtained from both workflows correlated to a significant degree. We found reasonable agreement of both workflows in protein fold changes between tested groups, resulting in unified lists of 20 and 22 proteins linked to oocyte maturity and blastocyst development, respectively. The Quad-Orbitrap workflow was best suited for an in-depth analysis without the need of extensive fractionation, especially of low abundant proteome, whereas the Triple Quad-TOF workflow allowed a more robust approach with a greater potential to increase in effectiveness with the growing number of analyzed samples after the initial effort of building a comprehensive spectral library.

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Cytokine antibody array profiling in human follicular fluid as a potential marker for oocyte quality

Fertility and Sterility ◽

10.1016/j.fertnstert.2016.07.554 ◽

2016 ◽

Vol 106 (3) ◽

pp. e187

Author(s):

M. Pavone ◽

J.M. Kelsh ◽

S. Malpani ◽

R. Confino ◽

S. Jasti ◽

...

Keyword(s):

Follicular Fluid ◽

Oocyte Quality ◽

Antibody Array ◽

Human Follicular Fluid ◽

Potential Marker ◽

Cytokine Antibody Array

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Oocyte Maturity and Human Follicular Fluid Prostanoids, Gonadotropins, and Prolactin After Administration of Clomiphene and Pergonal

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/jcem-65-3-402 ◽

1987 ◽

Vol 65 (3) ◽

pp. 402-406 ◽

Cited By ~ 17

Author(s):

J. Y. JEREMY ◽

F. E. OKONOFUA ◽

M. THOMAS ◽

J. WOJDYLA ◽

W. SMITH ◽

...

Keyword(s):

Follicular Fluid ◽

Oocyte Maturity ◽

Human Follicular Fluid

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Proteomic and peptidomic analysis of human follicular fluid by mass spectrometry as a potential tool for oocyte quality differentiation

10.26226/morressier.5912d9ead462b80292385fd5 ◽

2017 ◽

Author(s):

Joanna Liss

Keyword(s):

Mass Spectrometry ◽

Follicular Fluid ◽

Oocyte Quality ◽

Human Follicular Fluid ◽

Quality Differentiation ◽

Peptidomic Analysis ◽

Potential Tool

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Human Follicular Fluid Metabolomics Study of Follicular Development and Oocyte Quality

Chromatographia ◽

10.1007/s10337-017-3290-6 ◽

2017 ◽

Vol 80 (6) ◽

pp. 901-909 ◽

Cited By ~ 3

Author(s):

Zhengao Sun ◽

Haicui Wu ◽

Fang Lian ◽

Xingxing Zhang ◽

Conghui Pang ◽

...

Keyword(s):

Follicular Fluid ◽

Follicular Development ◽

Oocyte Quality ◽

Human Follicular Fluid ◽

Metabolomics Study

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Human follicular fluid levels of endothelins in relation to oocyte maturity status

Experimental and Clinical Endocrinology & Diabetes ◽

10.1055/s-0029-1211426 ◽

2009 ◽

Vol 104 (01) ◽

pp. 78-84 ◽

Cited By ~ 2

Author(s):

R. Sudik ◽

S. Chari ◽

E. Pascher ◽

G. Sturm

Keyword(s):

Follicular Fluid ◽

Oocyte Maturity ◽

Human Follicular Fluid ◽

Fluid Levels

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SWATHHM-Based Metabolomics of Follicular Fluid in Patients Shows That Progesterone Adversely Affects Oocyte Quality

BioMed Research International ◽

10.1155/2018/1780391 ◽

2018 ◽

Vol 2018 ◽

pp. 1-8 ◽

Cited By ~ 4

Author(s):

Zhengao Sun ◽

Jingyan Song ◽

Xingxing Zhang ◽

Aijuan Wang ◽

Ying Guo ◽

...

Keyword(s):

Follicular Fluid ◽

High Performance ◽

Principal Component ◽

Oocyte Quality ◽

Resolving Power ◽

Progesterone Level ◽

Control Group ◽

Human Follicular Fluid ◽

Spectrometric Data ◽

Flight Mass Spectrometry

Objective. We reveal the relationship between progesterone level in follicular fluid and oocyte quality based on sequential window acquisition of all theoretical fragment-ion spectra (SWATH™), a powerful high-resolution mass spectrometric data independent acquisition technique. Method. Follicular fluid samples were collected from 22 subjects (the level of progesterone > 1.5 ng/mL) of progesterone group, as well as from 22 subjects (the level of progesterone < 1.5 ng/mL) of control group, and analyzed using UPLC-Q-TOF. All methods were performed in accordance with ISO 9001:2008. Novel SWATH acquisition mode on an ultra-high performance liquid chromatography coupled with hybrid triple quadrupole time-of-flight mass spectrometry (with resolving power 20,000–40,000) was investigated for the analysis of human follicular fluid. The principal component variable grouping detects intersample variable correlation and groups variables with similar profiles which simplifies interpretation and highlights related ions and fragments. It can also extract product ion spectra from the data collected by fragmenting a wide precursor ion window. Results. Follicular fluid from the two groups differed with respect to five metabolites. Follicular fluid from the progesterone group contained elevated levels of 8-hydroxyguanosine and 4-hydroxynonenal and reduced levels of ATP, estradiol, and L-carnitine. The increased progesterone level on the day of HCG injection could negatively impact oocyte quality, thus reducing the pregnancy rate of IVF patients.

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Protein pathways working in human follicular fluid: the future for tailored IVF?

Expert Reviews in Molecular Medicine ◽

10.1017/erm.2016.4 ◽

2016 ◽

Vol 18 ◽

Cited By ~ 25

Author(s):

Laura Bianchi ◽

Assunta Gagliardi ◽

Claudia Landi ◽

Riccardo Focarelli ◽

Vincenzo De Leo ◽

...

Keyword(s):

Follicular Fluid ◽

Oocyte Quality ◽

In Vitro Fertilisation ◽

Protein Patterns ◽

Quality Estimation ◽

Human Follicular Fluid ◽

Physiological Processes ◽

Starting Point ◽

Pubmed Search

The human follicular fluid (HFF) contains molecules and proteins that may affect follicle growth, oocyte maturation and competence acquiring. Despite the numerous studies, an integrated broad overview on biomolecular and patho/physiological processes that are proved or supposed to take place in HFF during folliculogenesis and oocyte development is still missing. In this review we report, for the first time, all the proteins unambiguously detected in HFF and, applying DAVID (Database for Annotation, Visualization and Integrated Discovery) and MetaCore bioinformatic resources, we shed new lights on their functional correlation, delineating protein patterns and pathways with reasonable potentialities for oocyte quality estimation in in vitro fertilisation (IVF) programs. Performing a rigorous PubMed search, we redacted a list of 617 unique proteins unambiguously-annotated as HFF components. Their functional processing suggested the occurrence in HFF of a tight and highly dynamic functional-network, which is balanced by specific effectors, primarily involved in extracellular matrix degradation and remodelling, inflammation and coagulation. Metalloproteinases, thrombin and vitamin-D-receptor/retinoid-X-receptor-alpha resulted as the main key factors in the nets and their differential activity may be indicative of ovarian health and oocyte quality. Despite future accurate clinical investigations are absolutely needed, the present analysis may provide a starting point for more accurate oocyte quality estimation and for defining personalised therapies in reproductive medicine.

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