초록 ▼

Tyrosinase ELISA Kit 제작하기 위한 항체로서 4가지를 확보하였다. 그중
두가지 항체는 즉， Rabbit Anti-Mouse Tyrosinase Polyc1ona1 Antibody (Santa
Cruz Biotechnology, Cat No; sc-15341)와 Mouse Anti-Human Tyrosinase
Monoclonal Antibody (NeoMarkers, Cat. #MS-800-PIABX ) 는 상업적인 회사에 서
구입하였으며， 한 개는 상업적 회사 (고마바이오텍)에 의뢰하여 Mou

Tyrosinase ELISA Kit 제작하기 위한 항체로서 4가지를 확보하였다. 그중
두가지 항체는 즉， Rabbit Anti-Mouse Tyrosinase Polyc1ona1 Antibody (Santa
Cruz Biotechnology, Cat No; sc-15341)와 Mouse Anti-Human Tyrosinase
Monoclonal Antibody (NeoMarkers, Cat. #MS-800-PIABX ) 는 상업적인 회사에 서
구입하였으며， 한 개는 상업적 회사 (고마바이오텍)에 의뢰하여 Mouse
Anti-Human Tyrosinase를 제조하였고， 나머지 한 개는 본 연구실에서 직접 제조하
였다 ELISA는 측정 방식에 따라 (1) Antibody Sandwich ELISA 방법 과 (2)
lndirect Antibody ELISA로 구분할 수 있으며， 각각 장단점이 있다. 본 연구에서는
ELISA 제작에 필수적인 tyrosinase 에 대한 항체를 4 종류를 확립한 후 이를 이용
하여 Sandwich ELISA와 Indirect Antibody ELISA 제작하고， mouse melanoma
cell line에서 ELISA를 이용하여 tyrosinase 측정을 시도하였다.

Abstract ▼

Tyrosinase catalyses the rate-limiting step for the biosynthetic pathway of
melanin pigments. It is localized on melanomses in melanocytes. Although
various methods have been developed to measure the enzyme activities, it is not
easy to measure its activity accurately in the presence of inh

Tyrosinase catalyses the rate-limiting step for the biosynthetic pathway of
melanin pigments. It is localized on melanomses in melanocytes. Although
various methods have been developed to measure the enzyme activities, it is not
easy to measure its activity accurately in the presence of inhibitors in crude
samples. Therefore, it is helpful to measure the amount of tyrosinase in
melanocytes to understand the control of pigmentation. No quatitative analysis of
tyrosinase by immunologic method like ELISA have been established. Because
ELISA has several advantages: it used stable enzyme-conjugated reagents, has
a short reading time and no radioactive isotopes are necessary, we tried to
develop an ELISA for measuring mouse tyrosianse solubilized from B16 mouse
melanoma cells. A known amount of tyrosinase III solution was absorbed onto
micro titer plates and anti-tyrosinase antibodies were absorbed to tyrosinase.
Binding antibodies were detected by donkey anti-rabbit Ig or Sheep anti-mouse
Ig labeled with peroxidase. A sensitivity range of 1.25 ng to 10 ng/ well was
obtained. Although the level of sensitivity seems to be adequate to measure the
enzyme contnent in the mouse cells, tyrosinase extracted from B16 mouse
melanoma have not been detected by this indirect antibody ELISA. Because
most of tyrosinase is not in soluble form in melanoma cells, it should be
solubilized in order to measure its content using this assay.

목차 Contents

• 표지 ...1
• 최종보고서...3
• 제출문 ...6
• 요약문 ...7
• SUMMARY ...9
• 제1장 서론 ...10
• 제2장 국내,외 기술개발 현황 ...12
• 제3장 연구개발 수행내용 및 결과 ...14
• 가. ELISA 제작 원리 ...14
• 나. 연구 내용 ...14
• 제4장 연구개발 목표 달성도 및 대외 기여도 ...22
• 제5장 연구개발 결과의 활용성과 및 계획 ...23
• 1) Anti-tyrosinase antibody 제조 ...23
• 2) Indirect Antibody ELISA 제작 ...23
• 참고문헌 ...24