volume 44 issue 1 (march 2010) : 22 - 27
PRODUCTION OF MONOCLONAL ANTIBODY AGAINST MALE SPECIFIC ANTIGEN ON CELL MEMBRANE OF BOVINE SPERMATOZOA
1Graduated student of Graduate school, Chiang Mai University,
Chiang Mai, 50200, Thailand.
ABSTRACT
The objective of this study was to produce monoclonal antibody (MAb) against Y-bearing
spermatozoa. The antibody will be used in cytotoxic immunological reaction on the Y-bearing sperm,
which resulted in gender biased semen. Three BALB/c mice were immunized against dairy bull
spermatozoa and Freund’s complete adjuvant, 106 cells per 100 μl, at two week interval. The
spleenocytes, from the mouse that had produced antibodies to the Y-bearing spermatozoa, were
used for fusion with the myeloma cells line, X63-Ag8.653, with the aid of polyethylene glycol. It was
found, within 14-21 days after fusion, that 148 clones appeared. Three of those clones were capable
of binding specifically with white blood cells (WBC) from dairy bulls (n=5) and showed negative
response with the WBC from dairy heifers (n=5). The three antibodies were classified to be IgG. One
of them, clone number 1F9-3B10, was chosen due to its high antibody activity. The clone was tested
by using Immunofluorescent technique with WBC of 20 dairy bulls and 30 dairy heifers, the results
showed that 89.5 % of those bull WBC were recognized by the MAb from the clone number 1F9-3B10
(p005). In conclusion, the MAb tends to bind specifically with Y-bearing bull spermatozoa
spermatozoa. The antibody will be used in cytotoxic immunological reaction on the Y-bearing sperm,
which resulted in gender biased semen. Three BALB/c mice were immunized against dairy bull
spermatozoa and Freund’s complete adjuvant, 106 cells per 100 μl, at two week interval. The
spleenocytes, from the mouse that had produced antibodies to the Y-bearing spermatozoa, were
used for fusion with the myeloma cells line, X63-Ag8.653, with the aid of polyethylene glycol. It was
found, within 14-21 days after fusion, that 148 clones appeared. Three of those clones were capable
of binding specifically with white blood cells (WBC) from dairy bulls (n=5) and showed negative
response with the WBC from dairy heifers (n=5). The three antibodies were classified to be IgG. One
of them, clone number 1F9-3B10, was chosen due to its high antibody activity. The clone was tested
by using Immunofluorescent technique with WBC of 20 dairy bulls and 30 dairy heifers, the results
showed that 89.5 % of those bull WBC were recognized by the MAb from the clone number 1F9-3B10
(p005). In conclusion, the MAb tends to bind specifically with Y-bearing bull spermatozoa
REFERENCES
- Abbas A.K.(1994) General Properties of Immune Response in Cellular and Molecular Immunology. Saunders Co.,
- Pennsylvania.
- Amann R.P. (1990) Theriogenology 52 :1447-1457.
- Bennett, D. and Boyse, E.A. (1973) Nature 246 : 308 – 309.
- Booman P. et al(1989) Reproductive Immunology 15: 195-205.
- Borrebaeck C.A.K. and Moller, S.A. (1986) J. Immunology 136: 3710-3715.
- Bradley M.P. and Heslop, B.F. (1988) Human Genet 79: 347-351.
- Brunner, M. (1987) J. Reprod. Immunol. 11:181-191.
- Fu Q. et al (2007) Theriogenology 68 : 1211-1218.
- Goldberg E.J. (1971) Nature 232: 478-480.
- Hossepian de Lima, V.F.M. (1993) Theriogenology 39:1343-1352.
- Huhtinen M. et al (1997) Theriogenology 48: 361-367.
- Johnson L.J. et al (2005) Theriogenology 63: 615 – 624.
- Lyer S.V. (1989) Gamete Research 22: 37 – 49.
- Moce E. et al (2006) Theriogenology 66 :929-936.
- Morton K.M. et al (2006) Theriogenology 65: 1333 – 1345.
- Peter, A.T. et al (1993) Theriogenology 40:1177-1185.
- Ramalho T. et al (2004) Theriogenology 62: 1569-1576.
- Reilly B.D. and E.H. Goldberg. (1991) J. Immuno.Methods 142 : 121-126.
- Savikurki, H. et al (1981). J. Immuno. Methods. 41:343-349.
- Selden, J.R. (1982) Progr in Clinical Biolo Res 94:381-418.
- Seidel, G.E. (2007) Theriogenology 68: 443-446.
- Suh, T.K. et al (2007) Theriogenology 64: 1035-1048.
- Wachtel S. (1983) Fertility and Sterility 50: 355-360.
- Zaborski P. (1979) Transplantation 27 : 348-350
- Zavos, P.M. (1983) Theriogenology 20: 235-240
Disclaimer :
All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.
Copyright :
This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Published In
Indian Journal of Animal Research
Article Metrics
0
Views
Citations
Reviewed By
In this Article
APC
APC cover the cost of turning a manuscript into a published manuscript through peer-review process, editorial work as well as the cost of hosting, distributing, indexing and promoting the manuscript.
Publish With US
Submit your manuscript through user friendly platform and acquire the maximum impact for your research by publishing with ARCC Journals.
Become a Reviewer/Member
Join our esteemed reviewers panel and become an editorial board member with international experts in the domain of numerous specializations.
Open Access
Filling the gap between research and communication ARCC provide Open Access of all journals which empower research community in all the ways which is accessible to all.
Products and Services
We provide prime quality of services to assist you select right product of your requirement.
Support and Policies
Finest policies are designed to ensure world class support to our authors, members and readers. Our efficient team provides best possible support for you.
