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:: Volume 11, Issue 3 (Quarterly 2007) ::
Feyz 2007, 11(3): 24-30 Back to browse issues page
Purification of inhibin from human follicular fluid using monoclonal antibody
Mohammad Reza Sadeghi *, Mahshid Hodjat, Nima Naseri, Mahmoud Jeddi-Tehrani, Nariman Mosaffa, Mohammad Mahdi Akhondi, Roya Ghods, Ali Ahmad Bayat
, Sadeghi@avicenna.ac.ir
Abstract:   (8528 Views)

Background: Inhibin is a protein synthesized by granulosa and sertoli cells which preferentially inhibit pituitary secretion of FSH through a negative-feedback. Studies have showed that measurement of inhibin has a clinical role in understanding the fertility status of men and women and could also be used as a prognostic marker for pre-eclampsia and Down syndrome in fetus. There are many different multi-step procedures for the purification of inhibin. In this study we attempted to purify inhibin from human follicular fluid using immunoaffinity techniques.

Materials and Methods: Follicular fluid (FF) collected from women referring to Avicenna Infertility Clinic, was filtrated and subsequently concentrated by ammonium sulfate. The presence of inhibin in follicular fluid was detected using enzyme-linked immunosorbent assay . In order to purify inhibin, an affinity chromatography column using anti-inhibin monoclonal antibody was prepared. The purified protein was analyzed through SDS-PAGE and western blotting after a protein solution load of human inhibin into the column.

Results: The SDS-PAGE results of affinity proved the presence of inhibin following silver staining appeared as a single 32 kDa band . Western blotting revealed that specific anti-inhibin antibody was able to recognize inhibin epitopes. The present protocol for purification of inhibin is a technique with 92% yield.

Conclusion: This method is a sensitive procedure for high yielded productions of inhibin compared to the previously described methods, using HPLC and gel filtration. However, since preparing and obtaining follicular fluid, as a main source of inhibin, is rather difficult but suitable for laboratories, recombinant inhibin is recommended for mass production.

Keywords: Follicular fluid, Inhibin, Monoclonal antibody, Purification
Full-Text [PDF 249 kb]   (1888 Downloads)    
Type of Study: Research | Subject: General
Received: 2008/10/26
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Sadeghi M R, Hodjat M, Naseri N, Jeddi-Tehrani M, Mosaffa N, Akhondi M M, et al . Purification of inhibin from human follicular fluid using monoclonal antibody. Feyz. 2007; 11 (3) :24-30
URL: http://feyz.kaums.ac.ir/article-1-47-en.html


Volume 11, Issue 3 (Quarterly 2007) Back to browse issues page
مجله علمی پژوهشی فیض ::: دانشگاه علوم پزشکی کاشان KAUMS Journal ( FEYZ )
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