Capillary electrophoresis of human follicular fluid

Article

Wen, S., Perrett, D., Patel, P., Li, N., Docherty, S., Tozer, A. and Iles, R. 2009. Capillary electrophoresis of human follicular fluid. Journal of Chromatography B. 877 (31), pp. 3946-3952. https://doi.org/10.1016/j.jchromb.2009.09.046
TypeArticle
TitleCapillary electrophoresis of human follicular fluid
AuthorsWen, S., Perrett, D., Patel, P., Li, N., Docherty, S., Tozer, A. and Iles, R.
Abstract

Some of the major serum proteins that are also found in follicular fluid, including transferrin, α-macroglobulin and albumin, are thought to play a role in oocyte maturation. This study set out to identify proteins in human follicular fluid by capillary zone electrophoresis and to investigate their relationship to follicular/oocyte maturity and fertility outcome.
176 individual follicular fluid samples, from 30 women undertaking in vitro fertilization/ intracytoplasmic sperm injection (IVF/ICSI), were run using an optimized capillary zone electrophoresis method that gave a good separation of sixteen peaks in most samples. Nine of the peaks were identified and quantified but seven remain unknown and require further proteomic identification. Of the identified protein peaks, levels of each were corrected for follicular volume and total content calculated. No significant difference in protein levels was found with regard to oocyte recovery and fertilization. Protein concentrations tended to decrease as the follicular sphere increased whilst total content in follicular fluid increased in proportion to size. This is consistent with simple transudation across a sphere surface area which does not increase in proportion to the follicular fluid. This is not true of the concentration and content pattern of other proteins/biomolecules which are produced by follicular cells locally.
In conclusion, neither concentration nor absolute levels of nine major proteins identified in follicular fluids correlated with oocyte presence and fertility outcome. Future work to remove more concentrated proteins (e.g. albumin) would enhance separation of smaller peaks and identification of the unknown molecules.

Research GroupBiomarkers for Cancer group
PublisherElsevier
JournalJournal of Chromatography B
ISSN1570-0232
Publication dates
PrintDec 2009
Publication process dates
Deposited28 Oct 2009
Output statusPublished
Digital Object Identifier (DOI)https://doi.org/10.1016/j.jchromb.2009.09.046
LanguageEnglish
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Iles, R., Butler, S. and Jacoby, E. 1999. Dimerization of urinary beta-core/hCFbetacf: a cause of poor beta-core assay performance in Down syndrome screening studies. Prenatal diagnosis. 19 (8), pp. 790-792. https://doi.org/10.1002/(SICI)1097-0223(199908)19:8<790::AID-PD627>3.0.CO;2-Z
Urinary beta-core hCG: Screening for aneuploidies in early pregnancy (11-14 weeks’ gestation).
MacIntosh, M., Nicolaides, K., Noble, P., Chard, T., Gunn, L. and Iles, R. 1997. Urinary beta-core hCG: Screening for aneuploidies in early pregnancy (11-14 weeks’ gestation). Prenatal diagnosis. 17 (5), pp. 401-405. https://doi.org/10.1002/(SICI)1097-0223(199705)17:5<401::AID-PD40>3.0.CO;2-M
Substantial urinary concentrations of material resembling beta-core fragment of chorionic gonadotropin beta-subunit in mid-menstrual cycle
Neven, P., Iles, R., Howes, I., Sharma, K., Shepherd, J., Edwards, R., Collins, W. and Chard, T. 1993. Substantial urinary concentrations of material resembling beta-core fragment of chorionic gonadotropin beta-subunit in mid-menstrual cycle. Clinical Chemistry. 39 (9), pp. 1857-1860.
Production of placental alkaline phosphatase (PLAP) and PLAP-like material by epithelial germ cell and non-germ cell tumours in vitro
Iles, R., Ind, T. and Chard, T. 1994. Production of placental alkaline phosphatase (PLAP) and PLAP-like material by epithelial germ cell and non-germ cell tumours in vitro. British journal of cancer. 69 (2), pp. 274-278.
Diurnal variation of urinary "hCG beta subunit core fragment" production evaluated in patients with gynecological neoplasms
Neven, P., Iles, R., Carter, P., Shepherd, J. and Chard, T. 1994. Diurnal variation of urinary "hCG beta subunit core fragment" production evaluated in patients with gynecological neoplasms. Clinical Chemistry. 40 (3), pp. 484-485.
Characterisation of the differential expression of marker antigens by normal and malignant endometrial epithelium
Chatzaki, E., Gallagher, C., Iles, R., Ind, T., Nouri, A., Bax, C. and Grudzinskas, J. 1994. Characterisation of the differential expression of marker antigens by normal and malignant endometrial epithelium. British journal of cancer. 69 (6), pp. 1010-1014.
Serum placental-type alkaline phosphatase activity in women with squamous and glandular malignancies of the reproductive tract
Ind, T., Iles, R., Carter, P., Lowe, D., Shepherd, J., Hudson, C. and Chard, T. 1994. Serum placental-type alkaline phosphatase activity in women with squamous and glandular malignancies of the reproductive tract. Journal of Clinical Pathology. 47 (11), pp. 1035-1037.
Measurement of urinary beta core fragment of human chorionic gonadotrophin in women with vulvovaginal malignancy and its prognostic significance
Carter, P., Iles, R., Neven, P., Ind, T., Shepherd, J. and Chard, T. 1995. Measurement of urinary beta core fragment of human chorionic gonadotrophin in women with vulvovaginal malignancy and its prognostic significance. British journal of cancer. 71 (2), pp. 350-353.
The effects of beta-human chorionic gonadotrophin on the in vitro growth of bladder cancer cell lines
Gillott, D., Iles, R. and Chard, T. 1996. The effects of beta-human chorionic gonadotrophin on the in vitro growth of bladder cancer cell lines. British journal of cancer. 73 (3), pp. 323-326.
The temporary anatomical structures prominent in the first trimester may be fulfilling exchange functions assigned to the placenta in the second and third trimester
Docherty, S., Iles, R., Wathen, N. and Chard, T. 1996. The temporary anatomical structures prominent in the first trimester may be fulfilling exchange functions assigned to the placenta in the second and third trimester. Human Reproduction. 11 (6), pp. 1157-1161.
Prognostic significance of the bcl-2 apoptotic family of proteins in primary and recurrent cervical cancer
Crawford, R., Caldwell, C., Iles, R., Lowe, D., Shepherd, J. and Chard, T. 1998. Prognostic significance of the bcl-2 apoptotic family of proteins in primary and recurrent cervical cancer. British journal of cancer. 78 (2), pp. 210-214.
Concentrations of placental protein 14 in uterine flushings from infertile women: validation of the collection technique and method of expression of results
Hamilton, J., Iles, R., Gunn, L., Wilson, C., Lower, A., Chard, T. and Grudzinskas, J. 1998. Concentrations of placental protein 14 in uterine flushings from infertile women: validation of the collection technique and method of expression of results. Human Reproduction. 13 (12), pp. 3357-3362.
The prognostic significance of beta human chorionic gonadotrophin and its metabolites in women with cervical carcinoma
Crawford, R., Iles, R., Carter, P., Caldwell, C., Shepherd, J. and Chard, T. 1998. The prognostic significance of beta human chorionic gonadotrophin and its metabolites in women with cervical carcinoma. Journal of Clinical Pathology. 51 (9), pp. 685-688.
Cross-Reaction with luteinizing hormone ß-Core is responsible for the age-dependent increase of immunoreactive ß-Core fragment of human chorionic gonadotropin in women with nonmalignant conditions
Iles, R., Javid, M., Gunn, L. and Chard, T. 1999. Cross-Reaction with luteinizing hormone ß-Core is responsible for the age-dependent increase of immunoreactive ß-Core fragment of human chorionic gonadotropin in women with nonmalignant conditions. Clinical Chemistry. 45, pp. 532-538.
The beta-subunit of human chorionic gonadotrophin exists as a homodimer
Butler, S., Laidler, P., Porter, J., Kicman, A., Chard, T., Cowan, D. and Iles, R. 1999. The beta-subunit of human chorionic gonadotrophin exists as a homodimer. Journal of molecular endocrinology. 22, pp. 185-192.
Clinical and endocrinological changes after electro-acupuncture treatment in patients with osteoarthritis of the knee
Ahsin, S., Saleem, S., Bhatti, A., Aslam, M. and Iles, R. 2009. Clinical and endocrinological changes after electro-acupuncture treatment in patients with osteoarthritis of the knee. Pain. 147 (1-3), pp. 60-66. https://doi.org/10.1016/j.pain.2009.08.004
The role of glycosylation in regulating glycoprotein hormone free alpha subunit and free beta subunit combination in the extra-embryonic coelomic fluid of early pregnancy
Blithe, D. and Iles, R. 1995. The role of glycosylation in regulating glycoprotein hormone free alpha subunit and free beta subunit combination in the extra-embryonic coelomic fluid of early pregnancy. Endocrinology. 136 (3), pp. 903-910. https://doi.org/10.1210/en.136.3.903
Can biological markers improve the management of breast cancer patients?
Leake, R., Anderson, L., Dowsett, M., Iles, R., Nicholson, R., Robertson, J., Walker, R. and UKCCR Biomarker group 2000. Can biological markers improve the management of breast cancer patients? British journal of cancer. 82 (10), pp. 1625-1626. https://doi.org/10.1054/bjoc.2000.1159
Biological markers: maintaining standards
Leake, R., Anderson, L., Dowsett, M., Iles, R., Nicholson, R., Robertson, J., Walker, R. and UKCCCR Biomarker group 2000. Biological markers: maintaining standards. British journal of cancer. 82 (10), pp. 1627-1628. https://doi.org/10.1054/bjoc.2000.1158
Determination of the glycoforms of human chorionic gonadotropin b-core fragment by matrix-assisted laser desoption/ionisation time of flight mass spectrometry
Jacoby, E., Kicman, A., Laidler, P. and Iles, R. 2000. Determination of the glycoforms of human chorionic gonadotropin b-core fragment by matrix-assisted laser desoption/ionisation time of flight mass spectrometry. Clinical Chemistry. 46 (11), pp. 1796-1803.
Regulation of endometrial cancer cell growth by luteinising hormone (LH) and follicle stimulating hormone (FSH)
Davies, S., Bax, C., Chatzaki, E., Chard, T. and Iles, R. 2000. Regulation of endometrial cancer cell growth by luteinising hormone (LH) and follicle stimulating hormone (FSH). British journal of cancer. 83 (12), pp. 1730-1734. https://doi.org/10.1054/bjoc.2000.1507
The increase in bladder carcinoma cell population induced by the free beta subunit of human chorionic gonadotrophin is a result of an anti-apoptotic effect and not cell proliferation
Butler, S., Ikram, M., Mathieu, S. and Iles, R. 2000. The increase in bladder carcinoma cell population induced by the free beta subunit of human chorionic gonadotrophin is a result of an anti-apoptotic effect and not cell proliferation. British journal of cancer. 82 (9), pp. 1553-1556. https://doi.org/10.1054/bjoc.2000.1177
The analysis of human amniotic fluid using capillary electrophoresis.
Stewart, C., Iles, R. and Perrett, D. 2001. The analysis of human amniotic fluid using capillary electrophoresis. Electrophoresis. 22 (6), pp. 1136-1142. https://doi.org/10.1002/1522-2683()22:6<1136::AID-ELPS1136>3.0.CO;2-A
High concentrations of CA 125 in uterine flushings: influence of cause of infertility and menstrual cycle day.
Hamilton, J., Iles, R., Gunn, L., Wilson, C., Lower, A. and Grudzinskas, J. 2002. High concentrations of CA 125 in uterine flushings: influence of cause of infertility and menstrual cycle day. Gynecological endocrinology..
Reduction of bladder cancer cell growth in response to hCGb CTP37 vaccinated mouse serum.
Butler, S., Staite, E. and Iles, R. 2003. Reduction of bladder cancer cell growth in response to hCGb CTP37 vaccinated mouse serum. Oncology research. 14 (2), pp. 93-100.
Identification of post-translational modifications resulting from LH beta polymorphisms by matrix-assisted laser desorption time-of-flight mass spectrometric analysis of pituitary LH beta core fragment
Jacoby, E., Kicman, A. and Iles, R. 2003. Identification of post-translational modifications resulting from LH beta polymorphisms by matrix-assisted laser desorption time-of-flight mass spectrometric analysis of pituitary LH beta core fragment. Journal of molecular endocrinology. 30 (2), pp. 239-252. https://doi.org/10.1677/jme.0.0300239
The recently identified homodimeric beta-beta subunit of human chorionic gonadotrophin (hCGbb) is no more biologically active than the free beta subunit (hCGb)
Butler, S. and Iles, R. 2004. The recently identified homodimeric beta-beta subunit of human chorionic gonadotrophin (hCGbb) is no more biologically active than the free beta subunit (hCGb). Tumor Biology. 25 (1-2), pp. 18-23. https://doi.org/10.1159/000077719
Serum amino-terminal propeptide of type 1 procollagen (P1NP) in prostate cancer: a potential predictor of bone metastases and prognosticator for disease progression and survival.
Thurairaja, R., Iles, R., Jefferson, K., McFarlane, J. and Persad, R. 2006. Serum amino-terminal propeptide of type 1 procollagen (P1NP) in prostate cancer: a potential predictor of bone metastases and prognosticator for disease progression and survival. Urologia internationalis. 76 (1), pp. 67-71.
Immunoassay detection of human chorionic gonadotrophin beta (hCGb) expression by prostate cancer is of no clinical significance
Otite, U., Baithun, S., Chinegwundon, F., Nargund, V. and Iles, R. 2006. Immunoassay detection of human chorionic gonadotrophin beta (hCGb) expression by prostate cancer is of no clinical significance. Tumor Biology. 27, pp. 181-186.
Gonadotropins and prostate cancer: revisited.
Thwaini, A., Naase, M., Chinegwundon, F., Baithun, S., Ghali, L., Shergill, I. and Iles, R. 2006. Gonadotropins and prostate cancer: revisited. Urologia internationalis. 77 (4), pp. 289-296. https://doi.org/10.1159/000096330
Ectopic hCGbeta expression by epithelial cancer: malignant behaviour, metastasis and inhibition of tumor cell apoptosis
Iles, R. 2007. Ectopic hCGbeta expression by epithelial cancer: malignant behaviour, metastasis and inhibition of tumor cell apoptosis. Molecular and Cellular Endocrinology. 260-26, pp. 264-270. https://doi.org/10.1016/j.mce.2006.02.019
An introduction to mass spectrometry based proteomics-detection and characterization of gonadotropins and related molecules
Kicman, A., Parkin, M. and Iles, R. 2007. An introduction to mass spectrometry based proteomics-detection and characterization of gonadotropins and related molecules. Molecular and Cellular Endocrinology. 260-26, pp. 212-227. https://doi.org/10.1016/j.mce.2006.02.022
Towards the development of an electrochemical biosensor for hCGβ detection
Kassanos, P., Iles, R., Bayford, R. and Demosthenous, A. 2008. Towards the development of an electrochemical biosensor for hCGβ detection. Physiological Measurement. 29 (6), pp. S241-S254. https://doi.org/10.1088/0967-3334/29/6/S21
The role of glycoprotein hormones in endometrial cancer. (in Part 7: Hormones & hormone receptor)
Bax, C., Davies, S., Chatzaki, E., Butler, S. and Iles, R. 2003. The role of glycoprotein hormones in endometrial cancer. (in Part 7: Hormones & hormone receptor). in: Nishida, M. and Kuramoto, H. (ed.) Cell and molecular biology of endometrial carcinoma. London Springer. pp. 195-206
Molecular cell biology and genetic disorders
Iles, R. 2009. Molecular cell biology and genetic disorders. in: Kumar, P. and Clark, M. (ed.) Kumar & Clark’s clinical medicine. Edinburgh Elsevier Saunders. pp. 19-52
Chapter 1: Basic sciences: the cell.
Iles, R. 2008. Chapter 1: Basic sciences: the cell. in: Nargund, V., Raghavan, D. and Sandler, H. (ed.) Urological oncology. London Springer. pp. 3-36
Human chorionic gonadotropin isoforms in the diagnosis of ectopic pregnancy
Iles, R., Borrelli, P., Butler, S. and Staite, E. 2003. Human chorionic gonadotropin isoforms in the diagnosis of ectopic pregnancy. Clinical Chemistry. 49 (12), pp. 2045-2049. https://doi.org/10.1373/clinchem.2003.022095
The free monomeric beta subunit of human chorionic gonadotrophin (hCG beta) and the recently identified homodimeric beta-beta subunit (hCG beta beta) both have autocrine growth effects
Butler, S. and Iles, R. 2004. The free monomeric beta subunit of human chorionic gonadotrophin (hCG beta) and the recently identified homodimeric beta-beta subunit (hCG beta beta) both have autocrine growth effects. Tumor Biology. 25 (1-2), pp. 18-23. https://doi.org/10.1159/000077719
Enhanced affinity capture MALDI-TOF MS: orientation of an immunoglobulin G using recombinant protein G
Iles, R., Bansal, S., Jacoby, E., Neubert, H., Cowan, D. and Kicman, A. 2002. Enhanced affinity capture MALDI-TOF MS: orientation of an immunoglobulin G using recombinant protein G. Analytical Chemistry. 74 (15), pp. 3677-3683. https://doi.org/10.1021/ac025558z
Identification of placental transforming growth factor-beta and bikunin metabolites as contaminants of pharmaceutical human chorionic gonadotrophin preparations by proteomic techniques
Iles, R., Kicman, A., Malatos, S. and Neubert, H. 2005. Identification of placental transforming growth factor-beta and bikunin metabolites as contaminants of pharmaceutical human chorionic gonadotrophin preparations by proteomic techniques. Molecular and Cellular Proteomics. 4 (7), pp. 984-992. https://doi.org/10.1074/mcp.M500085-MCP200
Ectopic human chorionic gonadotropin beta secretion by epithelial tumors and human chorionic gonadotropin beta-induced apoptosis in Kaposi's sarcoma: is there a connection?
Iles, R. and Butler, S. 2003. Ectopic human chorionic gonadotropin beta secretion by epithelial tumors and human chorionic gonadotropin beta-induced apoptosis in Kaposi's sarcoma: is there a connection? Clinical Cancer Research. 9 (13), pp. 4666-4673.
Human granulosa-lutein cell in vitro production of progesterone, inhibin A, inhibin B, and activin A are dependent on follicular size and not the presence of the oocyte
Wen, S., Tozer, A., Li, D., Docherty, S., Al-Shawaf, T. and Iles, R. 2008. Human granulosa-lutein cell in vitro production of progesterone, inhibin A, inhibin B, and activin A are dependent on follicular size and not the presence of the oocyte. Fertility and Sterility. 89 (5), pp. 1406-1413. https://doi.org/10.1016/j.fertnstert.2007.03.086
Follicular fluid levels of inhibin A, inhibin B, and activin A levels reflect changes in follicle size but are not independent markers of the oocyte's ability to fertilize
Wen, S., Tozer, A., Butler, S., Bell, C., Docherty, S. and Iles, R. 2006. Follicular fluid levels of inhibin A, inhibin B, and activin A levels reflect changes in follicle size but are not independent markers of the oocyte's ability to fertilize. Fertility and Sterility. 85 (6), pp. 1723-1729. https://doi.org/10.1016/j.fertnstert.2005.11.058
The effects of 'coasting' on follicular fluid concentrations of vascular endothelial growth factor in women at risk of developing ovarian hyperstimulation syndrome
Tozer, A., Iles, R., Iammarrone, E., Gillott, C., Al-Shawaf, T. and Grudzinskas, J. 2004. The effects of 'coasting' on follicular fluid concentrations of vascular endothelial growth factor in women at risk of developing ovarian hyperstimulation syndrome. Human Reproduction. 19 (3), pp. 522-528. https://doi.org/10.1093/humrep/deh096
Characteristics of populations of granulosa cells from individual follicles in women undergoing 'coasting' during controlled ovarian stimulation (COS) for IVF
Tozer, A., Iles, R., Iammarrone, E., Gillott, C., Wen, S., Al-Shawaf, T. and Grudzinskas, J. 2004. Characteristics of populations of granulosa cells from individual follicles in women undergoing 'coasting' during controlled ovarian stimulation (COS) for IVF. Human Reproduction. 19 (11), pp. 2561-2568. https://doi.org/10.1093/humrep/deh487