PURIFICATION OF INHIBIN FROM HUMAN FOLLICULAR FLUID USING MONOCLONAL ANTIBODY

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SADEGHI M.R.; HOJAT MAHSHID; NASERI NIMA; JEDI TEHRANI MAHMOUD; MOSAFA NARIMAN; AKHOUNDI MOHAMMAD MAHDI; GHODS R.; BAYAT A.A.

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Journal: Feyz Medical Sciences Journal Year:2007 | Volume:11 | Issue:3 (SERIAL 43) Page(s): 24-30

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Title

PURIFICATION OF INHIBIN FROM HUMAN FOLLICULAR FLUID USING MONOCLONAL ANTIBODY

Author(s)

Keywords

FOLLICULAR FLUIDQ3

INHIBINQ3

MONOCLONAL ANTIBODYQ3

PURIFICATIONQ3

Abstract

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 antiinhibin 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.

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References

SURVEY OF SERUM INHIBIN B AS A RELIABLE MARKER FOR PREDICTION OF TESTICULAR SPERMATOZOA IN TESTES OF AZOOSPERMIC PATIENTS

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APA: Copy

SADEGHI, M.R., HOJAT, MAHSHID, NASERI, NIMA, JEDI TEHRANI, MAHMOUD, MOSAFA, NARIMAN, AKHOUNDI, MOHAMMAD MAHDI, GHODS, R., & BAYAT, A.A.. (2007). PURIFICATION OF INHIBIN FROM HUMAN FOLLICULAR FLUID USING MONOCLONAL ANTIBODY. FEYZ, 11(3 (SERIAL 43)), 24-30. SID. https://sid.ir/paper/40591/en

Vancouver: Copy

SADEGHI M.R., HOJAT MAHSHID, NASERI NIMA, JEDI TEHRANI MAHMOUD, MOSAFA NARIMAN, AKHOUNDI MOHAMMAD MAHDI, GHODS R., BAYAT A.A.. PURIFICATION OF INHIBIN FROM HUMAN FOLLICULAR FLUID USING MONOCLONAL ANTIBODY. FEYZ[Internet]. 2007;11(3 (SERIAL 43)):24-30. Available from: https://sid.ir/paper/40591/en

IEEE: Copy

M.R. SADEGHI, MAHSHID HOJAT, NIMA NASERI, MAHMOUD JEDI TEHRANI, NARIMAN MOSAFA, MOHAMMAD MAHDI AKHOUNDI, R. GHODS, and A.A. BAYAT, “PURIFICATION OF INHIBIN FROM HUMAN FOLLICULAR FLUID USING MONOCLONAL ANTIBODY,” FEYZ, vol. 11, no. 3 (SERIAL 43), pp. 24–30, 2007, [Online]. Available: https://sid.ir/paper/40591/en

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