| Development of Chemiluminescence Immunoassay(CIA) & ELISA for the Detection of Anti-sperm Antibodies in Male Serum |
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SC Kim1, KS Lee2, YK Kim2, CK Kim2, KH Choi2, OJ Kwon2, JB Kim2 |
1Department of Urology, College of Medicine, Chung-Ang University
2College of Animal Husbandry, Kon-Kuk University |
| 항정자 항체 검출을 위한 CIA 및 ELISA 개발을 위한 기초 연구 |
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김세철1, 이기순2, 김윤규2, 김창규2, 최경호2, 권오중2, 김종배2 |
1중앙대학교 의과대학 비뇨기과학교실
2건국대학교 축산대학 |
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| Abstract |
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New immunoassay systems for the detection of anti-sperm antibodies were developed. For this, sperm surface protein was purified by the immunoaffinity column prepared by the coupling of rabbit anti-human IgG antibodies to Sepharose-4B. Fraction eluted by tris-HCI buffer containing SDS showed a single band having molecular weight of about 60KD on electrophoresis. Enzyme HRP labelled goat anti-human IgG and chemiluminescence aminobutylethyl-isoluminol(ABEI) labelled rabbit anti-human IgG were used for ELISA and CIA, respectively. These two labelled conjugate bound well with human IgG. When serum dilution curves were made to titrate positive serums, two kinds of curves with steep and sluggish slopes were obtained Serum samples were categorized into 3 groups: positive, weak positive and negative based on slope of curve and O.D. values at 1:160 dilution of serum. When ELISA and CIA were compared to conventional method Kibrick test by the determinations of 62 male serums with different diagnosis, the results of ELISA and CIA agreed well, but both disagreed with that of Kibrick test. This study showed that purified sperm surface antigen can be used to develope solid-phase immunoassay systems such as ELISA and CIA which may eliminate the problems encounted the immobilization of living sperm in other tests. |
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