Hanrieder Jörg, Nyakas Adrien, Naessén Tord, Bergquist Jonas
Department of Physical and Analytical Chemistry, Analytical Chemistry, Uppsala University, Uppsala, Sweden.
J Proteome Res. 2008 Jan;7(1):443-9. doi: 10.1021/pr070277z. Epub 2007 Nov 30.
Human follicular fluid (hFF) is the in vivo environment of oocytes during follicular maturation in the ovaries. It contains a huge variety of compounds such as, e.g., proteins that might play an important role in follicular development and oocyte growth. Previous proteomic studies on follicular fluid have isolated and already identified a certain number of proteins. Nevertheless, only a small part of proteins present in follicular fluid have been covered so far and a large number have still not been identified. Therefore, the need for new, more resolving, and sensitive approaches in proteome research is evident. We utilized a proteomic setup based on in solution isoelectric focusing (IEF) and reversed-phase nanoliquid chromatography coupled to matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (nano-LC MALDI TOF/TOF MS) for in depth protein analysis of human follicular fluid samples of patients undergoing controlled ovarian hyper stimulation (COH) for in vitro fertilization therapy (IVF). This approach led to the significant identification of 69 proteins, where 32 have not been reported before to be found in human follicular fluid with proteomic methods. Among these findings, at least two relevant compounds essentially involved in hormone secretion regulation during the folliculogenetic process were identified: sex hormone binding globulin (SHBG) and inhibin A (INHA). To confirm these results, both proteins were further validated by immunoassays.
人卵泡液（hFF）是卵巢中卵泡成熟过程中卵母细胞的体内环境。它含有大量的化合物，例如可能在卵泡发育和卵母细胞生长中起重要作用的蛋白质。先前关于卵泡液的蛋白质组学研究已经分离并鉴定了一定数量的蛋白质。然而，到目前为止，卵泡液中存在的蛋白质只有一小部分被涵盖，大量蛋白质仍未被鉴定。因此，蛋白质组研究中需要新的、分辨率更高且更灵敏的方法是显而易见的。我们采用了一种基于溶液等电聚焦（IEF）和反相纳升液相色谱与基质辅助激光解吸/电离飞行时间串联质谱联用（nano-LC MALDI TOF/TOF MS）的蛋白质组学设置，对接受体外受精治疗（IVF）的控制性卵巢过度刺激（COH）患者的人卵泡液样本进行深度蛋白质分析。这种方法显著鉴定出69种蛋白质，其中32种以前尚未有蛋白质组学方法在人卵泡液中被报道过。在这些发现中，鉴定出至少两种在卵泡发生过程中基本参与激素分泌调节的相关化合物：性激素结合球蛋白（SHBG）和抑制素A（INHA）。为了证实这些结果，通过免疫测定对这两种蛋白质进行了进一步验证。
