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New fertility diagnostic kits

Preparation of new monoclonal antibodies against human
sperm proteins. In vitro production of monoclonal
antibodies, development of diagnostic kits for human sperm
pathology and assisted reproduction.

The infertility of human population is already a problem and will be even more for future generations. Recent studies suggest that over 15% of married couples suffer fertility problems. The male factor is the cause of infertility in more than 40% of problematic couples. However little is known about the pathophysiology of such sperm. The aim of the present project is to prepare methods, tools and kits for clinical diagnosis of pathologic human spermatozoa and to apply them in assisted reproduction. The first step of the project will be to prepare new monoclonal antibodies against human cell surface (including spermadhesins) and intra-acrosome sperm proteins reacting on acetone fixed smears of sperm and also on formaldehyde-fixed paraffin sections of reproductive organs. Cell surface sperm proteins are important in the process of primary binding of sperms to oocytes (1,2,3), intra-acrosomal proteins play a role as secondary binding proteins after acrosome reaction (4,5). Zona pellucida sperm binding surface proteins and intra-acrosomal proteins are presented in normospermic sperm cells and ensure the binding of sperm to oocyte and its penetration. Monoclonal antibodies can be used for the detection of zona pellucida sperm binding proteins (4) and for studying changes in the acrosomal status of sperm, integrity of cell membrane and presence of intra-acrosomal proteins (6,7). Acrosome membrane integrity is essential for the presence of proteolytic enzymes inside the acrosome, a component necessary for penetration of sperm into oocyte. Spermatozoa from subjects with a low amount of certain surface proteins are associated with certain cases of idiopathic infertility (8). Fragile or damaged spermatozoa after spontaneous acrosome reaction (9) are also connected with problems of defective binding of sperm to oocytes resulting probably in fertility defects. New diagnostic kits based on these new antibodies will therefore be developed permitting their diagnostic application for the detection of pathologic spermatozoa. One diagnostic kit will be utilized for detection of selected cell surface proteins and another one for detection of intra-acrosomal proteins. Kits may also be used for selection of suitable spermatozoa in assisted reproduction. The second aim of the project is to find relation between the ability of sperm to fertilize eggs and tyrosine phosphorylation (TP) of sperm proteins. TP is conserved among all mammalian species (10,11,12). We intend to establish the correlation between the level of TP and sperm pathology and will attempt to find out the appropriate combination of activators and inhibitors of phosphorylation to bring the phosphorylation back to the levels seen in normal sperm. The close contact with clinics in all participating countries will provide unique sperm samples for preparation of hybridoma cells and for subsequent testing of the diagnostic value of selected monoclonal antibodies and method of phosphorylation. 1. Ward CR, Kopf GS: Dev Biol 158:9-34,1993 2. Yanagimachi R: Physiol Reprod pp, 81-182,1994 3. Calvete JJ, Ensslin M, Mburu J, Iborra A, Martinez P, Adermann K, Waberski D, Sanz L, Topfer-Petersen E, Weitze KF, Einarsson S. Rodriquez Martinez H: Biol Reprod 57: 735-742,1997 4. Moos J, Peknicova J, Tesarik J: Biochim Biophys Acta 1176:199-207,1993 5. Brucker C, Lipfors GB: Hum Reprod 1:56-62,1995 6. Peknicova J, Moos J, Mollova M, Srsen V, Capkova J: Anim Reprod Sci 35:255-271,1994 7. Geussova G, Peknicova J, Capkova J, Kalab P, Moos J, Philimonenko VV, Hozak P: Andrologia 29:261-268,1997 8. Boue F, Sullivan R: Biol Reprod 54:1018-1024,1996 9. Peknicova J., Moos J.: Fol Biol: 44:93-96,1998 10. Leyton L, Saling P: J Cell Biol 108: 2163, 1989 11. Tesarik J, Moos J, Mendoza C: Endocrinology 133:328,1993 12. Morte C, Ibbora A, Martinez P: Molec Reprod Dev 50:113-120,1998. Keywords: fertility, diagnosis, monoclonal antibodies.
Acronym: 
NFDK-MOAB
Project ID: 
1 985
Start date: 
01-01-1999
Project Duration: 
60months
Project costs: 
870 000.00€
Technological Area: 
Market Area: 

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