Lactoferrin은 철과 결합하는 당단백질로 주로 모유나 우유, 혈액, 침 눈물, 점액분비물 등에 존재하고 있다. 최초의 Lactoferrin의 소의 유즙으로부터 정제, 동정되었고, 곧 쥐나 염소, 토끼, 개, 사람 등 여러 포유동물의 유즙에서도 차례로 분리, 정제되었다. 이미 보고된 바와 마찬가지로, Lactoferrin은 항균작용을 비롯해 여러 가지 생물학적 기능과 면역조절작용, 항암능력을 갖고 있다. 본 연구에서는 이러한 생리활성기능을 가진 Lactoferrin과 ...
Lactoferrin은 철과 결합하는 당단백질로 주로 모유나 우유, 혈액, 침 눈물, 점액분비물 등에 존재하고 있다. 최초의 Lactoferrin의 소의 유즙으로부터 정제, 동정되었고, 곧 쥐나 염소, 토끼, 개, 사람 등 여러 포유동물의 유즙에서도 차례로 분리, 정제되었다. 이미 보고된 바와 마찬가지로, Lactoferrin은 항균작용을 비롯해 여러 가지 생물학적 기능과 면역조절작용, 항암능력을 갖고 있다. 본 연구에서는 이러한 생리활성기능을 가진 Lactoferrin과 유청단백질을 산양의 초유에서 분리해내 사람의 주요 암세포종 (colorectal cancer cell HT-29, lung cancer cell A549, gastric cancer cell KATO-III, uterus cancer cell HeLa, and breast cancer cell ZR-75-1) 에 대해 in vitro 실험을 시행하였고, 이를 우유의 Lactoferrin 및 유청단백질과 비교하여 그 항암능을 비교연구하였다. 산양초유로부터 지방을 제거하고 산처리를 이용해 casein을 제거한 후 UF-membrane filtering을 통해 whey protein을 농축시켰다. 농축된 whey는 ion-exchange chromatography (CM-SephadexC-50-12)를 이용해 lactoferrin crude fraction을 얻었고, affinity chromatography (Hi-TrapTM Heparin HP)로 고순도의 lactoferrin을 분리하였다. Lactoferrin의 순도측정은 전기영동법을 이용하였는데 AF-column을 통과하여 얻은 fractions에서는 lactoferrin단일밴드를 확인할 수 있었다. 분리된 산양유 whey와 lactoferrin은 우유의 whey와 lactoferrin과 함께 항암실험에 사용되었다. MTT법을 이용하여 다양한 농도의 유청단백질과 lactoferrin을 처리한 후 세포주의 성장을 측정한 결과, 세포주의 종류에 따라 특이적인 성장 억제 효과가 관찰되었고, 특히 lactoferrin에서 더 높은 성장억제효과가 나타났다.
Lactoferrin은 철과 결합하는 당단백질로 주로 모유나 우유, 혈액, 침 눈물, 점액분비물 등에 존재하고 있다. 최초의 Lactoferrin의 소의 유즙으로부터 정제, 동정되었고, 곧 쥐나 염소, 토끼, 개, 사람 등 여러 포유동물의 유즙에서도 차례로 분리, 정제되었다. 이미 보고된 바와 마찬가지로, Lactoferrin은 항균작용을 비롯해 여러 가지 생물학적 기능과 면역조절작용, 항암능력을 갖고 있다. 본 연구에서는 이러한 생리활성기능을 가진 Lactoferrin과 유청단백질을 산양의 초유에서 분리해내 사람의 주요 암세포종 (colorectal cancer cell HT-29, lung cancer cell A549, gastric cancer cell KATO-III, uterus cancer cell HeLa, and breast cancer cell ZR-75-1) 에 대해 in vitro 실험을 시행하였고, 이를 우유의 Lactoferrin 및 유청단백질과 비교하여 그 항암능을 비교연구하였다. 산양초유로부터 지방을 제거하고 산처리를 이용해 casein을 제거한 후 UF-membrane filtering을 통해 whey protein을 농축시켰다. 농축된 whey는 ion-exchange chromatography (CM-SephadexC-50-12)를 이용해 lactoferrin crude fraction을 얻었고, affinity chromatography (Hi-TrapTM Heparin HP)로 고순도의 lactoferrin을 분리하였다. Lactoferrin의 순도측정은 전기영동법을 이용하였는데 AF-column을 통과하여 얻은 fractions에서는 lactoferrin단일밴드를 확인할 수 있었다. 분리된 산양유 whey와 lactoferrin은 우유의 whey와 lactoferrin과 함께 항암실험에 사용되었다. MTT법을 이용하여 다양한 농도의 유청단백질과 lactoferrin을 처리한 후 세포주의 성장을 측정한 결과, 세포주의 종류에 따라 특이적인 성장 억제 효과가 관찰되었고, 특히 lactoferrin에서 더 높은 성장억제효과가 나타났다.
Lactoferrin is an iron-binding glycoprotein secreted from mammary glands. It was well known that lactoferrin has a variety of biological activities such as immuno-modulation, anticancer activity. In this study lactoferrin was isolated from the goat colostrums and demonstrated the effect of lactoferr...
Lactoferrin is an iron-binding glycoprotein secreted from mammary glands. It was well known that lactoferrin has a variety of biological activities such as immuno-modulation, anticancer activity. In this study lactoferrin was isolated from the goat colostrums and demonstrated the effect of lactoferrin addition on the human colorectal cancer cell line HT-29, the human lung cancer cell line A549, the human gastric cancer cell line KATO-III, the human uterus cancer cell line HeLa, and the human breast cancer cell line ZR-75-1. From goat milk colostrums, the fat was skimmed and whey was separated by acid treatment. The goat lactoferrin was isolated by ion-exchange chromatography using CM-SephadexC-50-12 followed by affinity chromato- graphy on Hi-TrapTM Heparin HP. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis suggested the molecular mass of Saanen goat lactoferrin is 80 kDa. After purification of lactoferrin, the Goat whey and lactoferrin fraction which we purified, and bovine whey and lactoferrin was added to five kinds of human cancer cell lines. The cell proliferation inhibitory activity and cell viability was determined by MTT assay, and the concentration for 50% growth inhibition value are expressed on a molar and ㎍/ml basis. Various concentrations of goat and bovine milk protein components (whey and lactoferrin) treatment inhibited the growth inhibit- ory effect showed specificity depending on different cell lines. The each species lactoferrin were generally more active against human cancer cell lines than that of whey protein of each species. The bovine lactoferrin was the most active through the five human cancer cells and toxicity against five cancer cells was decreased goat lactoferrin, bovine whey protein, and goat whey protein in orderly. On the other hand bovine lactoferrin was the most effective against the HT-29, goat lactoferrin proved to be most effective against ZR-75-1.
Lactoferrin is an iron-binding glycoprotein secreted from mammary glands. It was well known that lactoferrin has a variety of biological activities such as immuno-modulation, anticancer activity. In this study lactoferrin was isolated from the goat colostrums and demonstrated the effect of lactoferrin addition on the human colorectal cancer cell line HT-29, the human lung cancer cell line A549, the human gastric cancer cell line KATO-III, the human uterus cancer cell line HeLa, and the human breast cancer cell line ZR-75-1. From goat milk colostrums, the fat was skimmed and whey was separated by acid treatment. The goat lactoferrin was isolated by ion-exchange chromatography using CM-SephadexC-50-12 followed by affinity chromato- graphy on Hi-TrapTM Heparin HP. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis suggested the molecular mass of Saanen goat lactoferrin is 80 kDa. After purification of lactoferrin, the Goat whey and lactoferrin fraction which we purified, and bovine whey and lactoferrin was added to five kinds of human cancer cell lines. The cell proliferation inhibitory activity and cell viability was determined by MTT assay, and the concentration for 50% growth inhibition value are expressed on a molar and ㎍/ml basis. Various concentrations of goat and bovine milk protein components (whey and lactoferrin) treatment inhibited the growth inhibit- ory effect showed specificity depending on different cell lines. The each species lactoferrin were generally more active against human cancer cell lines than that of whey protein of each species. The bovine lactoferrin was the most active through the five human cancer cells and toxicity against five cancer cells was decreased goat lactoferrin, bovine whey protein, and goat whey protein in orderly. On the other hand bovine lactoferrin was the most effective against the HT-29, goat lactoferrin proved to be most effective against ZR-75-1.
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