카드뮴폭로에 대한 생물학적 지표
Competitive ELISA에 의한 Metallothionein 정량Biological Index for Cadmium Exposure : Assay of Metallothionein by Competitive ELISA
- Authors
- 박정덕; 이우석; 공윤; 홍연표; 장임원
- Issue Date
- Sep-1996
- Publisher
- 대한산업의학회
- Keywords
- Metallothionein (MT); Cadmium Toxicity; Competitive ELISA, anti-MT-I
- Citation
- 대한산업의학회지, v.8, no.2, pp 258 - 271
- Pages
- 14
- Journal Title
- 대한산업의학회지
- Volume
- 8
- Number
- 2
- Start Page
- 258
- End Page
- 271
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/66619
- Abstract
- Metallothionein (MT) is a cadmium binding protein that played major roles in protective mechanism for cadmium toxicity. In the present study, competitive ELISA was estab- lished to assay the MT expression utilizing monospecific antibody which was generated against MT-I. A total of 10 Sprague-Dawley rats was injected with CdCla for two weeks to induce MT. The cytosolic fraction of rat liver was obtained by differential centrifuga- tion. Two major MT isozymes (MT-I & MT-II) at ca. 10 kDa were purified by Sephadex G-75 gel filtration followed by DEAE-Trisacryl-M anion exchange column chromatogra- phy, respectively. Two rabbits were immunized 4 times consecutively with the conjugate of purified MT-I. The sera were collected by heart puncture. When immunoblot. was car ried out, the immunized rabbit sera (anti-MT-1) exhibited specific immunoreactive band at MT-I while showed any cross reactions neither with MT-II nor with other cytosolic pro teins. By chequerboard titration using the monospecific antibody, the competitive ELISA was established. The dose-dependent relationship was observed between anti-MT-I anti- body and the amount of MT in biological samples (r=0.9980). These results suggested strongly that competitive ELISA is a simple, rapid and reproducible method for screening cadmium exposure.
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