A development plan for the immunodiagnostic kit of endometriosis

The aetiology of endometriosis is not well understood. Retrograde menstruation with implantation of endometrial fragments outside of uterus is the most widely accepted explanation for endometriosis. This commonly found disease is the main cause of infertility in women at reproductive age. However, there is no efficient and low invasive procedure for readily diagnosis of endometriosis, unless through the invasive procedure, laparoscopy or laparotomy to be confirmed. Currently, none of the reliable diagnostic markers for endometriosis is available. We have previously used proteomic approach to identify several serum proteins that may associate with the development of endometriosis. Based on the previous study, in 2008, the first US patent in a title of Diagnosis Method of Endometriosis by Detecting Biochemical Markers and Usage of These Biochemical Markers, W-C Yang, C-R Tzeng, and H-J Wang , US7399598, 2008/2.19~2025/10/14 was approved. Subsequently, the ROC patent was also approved (I 305837, 2009/2/1 – 2025/11/24. Republic of China). In addition, in 2009, the above invention was awarded as a Golden Award from 2009 National Invention and Creation Award from economic department. Since more new evidences wer found and 2 monoclonol antibodies against alpha 1 anti-trypson are produced and characterized, another US provisional patent for the monoclonal antibodies are filed for prepaing the application of PCT in order to apply the patent of other countries around the world. Our research team aims to develop the first generation of serum diagnostic kit for endometriosis in and achieve the technology transfer to collaborate with industry for the kit development. After screening the serum samples from 201 subjects (2008.2.1 -2008.12.15 and will have more samples to be collected after), we found 3 alpha 1-antitrypsin (A1AT) isoforms were significantly increased in sera of patients with endometriosis compared to the control of women without endometriosis (p<0.01). The major A1AT isoforms are 58 and 72 Da. After enzymatic analysis by digestion with N-glycanse, it appears that both 58 aand 72 kDa A1AT were glycosylated and sialyl Lewis X (sLeX), a glycoepitope is included. We therefore prepared the antibodies against 58 aand 72 kDa A1AT isoforms and the antibody against sLeX. The preliminary study showed that our homemade anti-A1AT antiserum can detect serum A1AT with high sensitivity. With 1:100000 dilution, the antiserum remained to distinguish A1AT in sera of patients with endometriosis whereas the commercially acailable monoclonal antibody lost its detection activity at 1:50000 dilution. In the continuous study, we will complete the antibody preparation. The sensitivity and specificity of the antibodies for screening serum of patients with endometriosis will be determined. In addition, the first generation of serum diagnostic kit for endometriosis will be designed and complete. The sensitivity and specificity of the diagnostic kit for endometriosis will be also evaluated. This study aims to develop a low invasive, low cost procedure for diagnosis of endometriosis. Early diagnosis and treatment of endometriosis will prevent the occurrence of endometriosis associated infertility and pregnancy loss.