Sperm preparation affects reactivity of monoclonal antibodies

Gupta, Satish K. ; Fulgham, David L. ; Alexander, Nancy J. (1990) Sperm preparation affects reactivity of monoclonal antibodies Journal of Reproductive Immunology, 18 (2). pp. 187-197. ISSN 0165-0378

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Official URL: http://www.sciencedirect.com/science/article/pii/0...

Related URL: http://dx.doi.org/10.1016/0165-0378(90)90016-Y


The polyacrylamide gel electrophoresis (SDS-PAGE) patterns of extracts prepared from thrice-washed human sperm were compared with extracts of swim-up and non-rise sperm. For sperm extraction sodium deoxycholate (DOC), octylphenoxypolyethoxyethanol (NP-40), dithiothreitol (DTT), lithium 3,5-diiodosalicylate (LIS), or 3-[(3-cholamidopropyl) dimethylammonio]-1-propane sulfonate (CHAPS) were used. A pellet of pooled thrice-washed semen was divided and extracted with these detergents. Similar gel electrophoresis profiles, as revealed by silver staining, were observed with the LIS, DOC, NP-40 and CHAPS extracts, whereas DTT extracts lacked the 150 and 200 kDa bands. Different thrice-washed semen pools varied in their protein pattern, probably due to the presence of sloughed germinal cells, white blood cells and sloughed cells from the reproductive tract. Swim-up sperm extracts had a different protein profile from that of thrice-washed pools but pools of swim-up sperm had a more constant protein pattern than those of thrice-washed semen pellets. Monoclonal antisperm antibodies did not always recognize antigens of the same molecular weights in western blots when extracts of swim-up sperm versus thrice-washed sperm were compared. In immunoblot studies with monoclonals (produced against thrice-washed sperm, cloned five times, reactive by ELISA and sperm function tests), reacted with 0.3% NP-40 extracts of thrice-washed sperm and with 0.3% NP-40 extract of swim-up sperm. These findings suggest that definition of antigens by monoclonal antisperm antibodies will be influenced by the nature of sperm extract employed.

Item Type:Article
Source:Copyright of this article belongs to Elsevier Science.
Keywords:Human Sperm; SDS-PAGE Electrophoresis; Immunoblotting; Monoclonal Antibodies
ID Code:72164
Deposited On:28 Nov 2011 06:07
Last Modified:28 Nov 2011 06:07

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