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구현정(Hyun-Jung Koo),하창우(Chang-Woo Ha),김성혁(Sung-Hyeok Kim),장소희(So-Hee Jang),임효선(Hyo-Sun Lim),김윤규(Youn-Kyu Kim),손은화(Eun-Hwa Sohn) 한국산학기술학회 2021 한국산학기술학회논문지 Vol.22 No.11
노근(Phragmitis rhizoma )은 갈대의 뿌리줄기를 말린 한약재로 열을 내리고 진액을 북돋는 청열약으로 알려져있다. 본 연구는 노근을 피부에 적용하기 위하여 안전한 추출조건에서 미백, 주름개선 및 보습효능에 대한 효과 기전을 규명하고 기능성 화장품 소재로서의 개발 가능성을 제시하고자 하였다. 70 % EtOH로 추출한 노근추출물을 이용하여 피부층을 이루는 멜라닌형성세포, 진피섬유세포, 각질형성세포에 미치는 세포독성을 측정하였다. 노근추출물의 미백, 주름개선, 보습에 대한 효능성을 평가하고, 노근의 지표성분 p -coumaric acid 함량을 분석하였다. 실험결과 노근추출물은 236±0.2 mg/kg의 p -coumaric acid를 함유하였으며, 200 μg/ml 이하의 농도에서 멜라닌형성세포, 진피섬유아세포, 각질형성세포 모두의 피부구성세포에 독성을 나타내지 않았다. 미백, 주름개선, 보습의 효능성 평가 부분에서 노근추출물 25와 50 μg/ml은 14.24 %, 17.52 %로 멜라닌 생성을 억제하는 미백효과를 나타내었으며(p <0.01), 진피섬유아세포의 콜라겐 분해효소 matrix metalloproteinase-1의 mRNA 발현과(p <0.01), 엘라스타제 활성을 유의성 있게 감소시킴으로서 주름개선 효과를 나타내었다(p <0.05). 또한, 노근추출물은 자외선B 노출에 의해 감소된 각질형성세포의 보습인자 hyaluronic acid synthase-2의 발현을 크게 회복하여 증가시킴으로써 보습효과를 보여주었다(p <0.01). 이와 같은 결과는 70 % EtOH로 추출한 노근추출물이 안전한 범위에서 미백, 주름개선, 보습 효과의 기능성 화장품 원료로 개발될 수 있다는 가능성을 제시한다. Nogeun (Phragmitis rhizoma ) is a dried herbal medicine derived from the rhizomes of reeds. The present study was aimed at investigating the whitening, anti-wrinkle, and moisturizing effects of P. rhizoma extracts and their safety aspects and to suggest the potential for its development as a functional cosmetic material. The study results showed that the 70 % ethyl alcohol (EtOH) extract of P. rhizoma contained 236±0.2 mg/kg of p -coumaric acid, and exhibited no cytotoxic effect on all the skin cells tested including melanocytes, dermal fibroblasts, and keratinocytes at concentrations at or below 200 μg/ml. In the evaluation of whitening, anti-wrinkle, and moisturizing effects, 25 and 50 μg/ml of P. rhizoma inhibited melanin production by 14.24 % and 17.52 %, respectively (p <0.01). P. rhizoma also reduced the mRNA expression of matrix metalloproteinase-1 and collagenase and decreased the activity of elastase in dermal fibroblasts. Additionally, P. rhizoma significantly increased the reduction of hyaluronic acid synthase-2, a moisturizing factor induced by UVB exposure in keratinocytes. These results suggest that the 70 % EtOH extract of P. rhizoma can be developed as a functional cosmetic material due to its skin whitening, anti-wrinkle, and moisturizing properties.
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김종배,김창규,최경호,김윤규 건국대학교 동물자원연구센터 1991 動物資源硏究誌 Vol.16 No.-
The double antibody, or second antibody, procedure is the most widely used system for separation of antibody bound and free fractions in immunoassay methods. The application of the method has ranged across the entire field of endocrine investigation from the assay of low molecular weight steroid hormones to the high molecular weight polypeptide and protein hormone. Second antibody was produced by immunizing a korean native goat with immunoglobulin<IgG> purified from rabbit. Then in order to develope 2nd-Ab as universal agents, first HRP was labeled to 2nd-Ab. Indirect ELISA was performed for soy protein . Second, the usefulness of goat anti-rabbit IgG antiserum for separation system was tested in the RIA for cortisol using anti-cortisol rabbit primary antisera.
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Hepatitis B virus preS1에 대한 생쥐 단일클론항체의 제조와 특성연구
김윤규,김희선,류춘제,진병래,홍효정 大韓免疫學會 1996 大韓免疫學會誌 Vol.18 No.3
To generate a murine monoclonal antibody (mAb) to the preSi antigen of hepatitis B virus (HBV), preSi fusion proteins maltose binding protein (MBP)-preSl and glutathione S-transferase (GST)-preSl/preS2 were expressed in Escherichia coli in a soluble form and purified from the cell lysate. Each fusion protein was immunized into BALB/c mice and the resulting antisera were titrated for the antibody to the preSi antigen. The result showed that the immunogenicity of the MBP-preSi was more effective than that of the GST preSi/preS2. Six hybridoma clones secreting good amount of the mAb were isolated and the isotype of each mAb was determined, which showed that two were IgGl and four were IgM. Among them, 1B3 antibody (IgGl), derived from the mice immunized with the MBP-preSi, was purified from the culture supernatant of the hybridoma and characterized. The purified 1B3 bound to the preSi in a dose-dependent manner and immunoprecipitated HBV particles as almost efficiently as anti-S and anti-preS2 mAbs which were previously shown to have the in vitro neutralizing activity for HBV. These results indicate that the MBP-preSl fusion protein can be an useful immunogen for the generation of the anti-preSi murine mAb and the 1B3 mAb may be useful to the fundamental and applied researches related with HBV.
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혈청 내의 총 Triiodothyronine 측정을 위한 경쟁적 효소면역분석법 개발
김종배,김창규,김윤규,박명례 대한내분비학회 1996 Endocrinology and metabolism Vol.11 No.4
Background: Triiodothyronine(T3) is a hormone secreted from thyroid gland which exerts a stimulating effect on metabolism. The disorder of thyroid system brings about several serious diseases like hypothymidism or hyperthyroidism. Therefore, the determination of T in blood is very important on monitoring thyroid function. Methods: Rabbit anti-T3 antibody was generated by immunization of T-BSA as an immunogen and purified hom antisera using Affi-gel protein A kit. The titer and specificity of purified antibody were characterized. To detect T3, competitive ELISA was performed using anti-T3 antibody and T3-HRP conjugate which was synthesized by glutaraldehyde method. The sensitivity and precision assay wer~e deterrnined and compared with that of RIA. Results: The titer of purified anti-T3 antibody was about 1:100 and the optimal dilution of T3- HRP conjugate was 1:1000. When the standard curve was constructed by ELISA, its sensitivity was about 0.5ng/ml. The eoefficient variations of intra- and inter-assay were 4.9~9.3% and 7.5~13.8%, respectively. The results obtained by ELISA and RIA correlated well with each other(n =50, r= 0.97), The linear regression equation was y= 1.09*0.08(P$lt;0.01). Conclusion: We successfully developed a method for the measurement of T3 on ELISA which was based on competitive reaction between antigen(T3) and enzyme labeled antigen(T3-HRP). These results demonstrated that competitive ELISA is a convenient, fast, reproducible and aecurate method for the determinstion of T in serum and can be used as practical alternative to RIA (J Kor Soc Endocrinol 11:447~454, 1996).
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