http://chineseinput.net/에서 pinyin(병음)방식으로 중국어를 변환할 수 있습니다.
변환된 중국어를 복사하여 사용하시면 됩니다.
김하형(Ha Hyung Kim),이광표(Kwang Pyo Lee),Koichi Kato,Yoji Arata 대한약학회 1995 약학회지 Vol.39 No.5
The anti-dansyl antibodies were specifically labeled with stable isotope by growing hybridoma cells in serum-free medium. Assignments of the observed carbonyl carbon resonances have been determined by using 13C-15N double labeling method in order to assign the Leu resonances. However, when the identical dipeptide appears more than twice in the polypeptide sequences, we applied the proteolytic fragments in the fragment-specific method. Carboxypeptidase B-treated antibody has also been used to assign the Lys-447 in C terminal amino acid. These unambiguously assigned carbonyl carbon resonances in antibodies are thought to be useful in elucidating not only the structure of antibodies but also the structure-function relationship in the antibody by 13C neuclear magnetic resonance spectroscopy.
NMR에 의한 anti-Ex-A IgG의 항원결합부위 해석
김하형(Ha Hyung Kim),이광표(Kwang Pyo Lee),가토 코이치(Koichi Kato),아라타 요우지(Yoji Arata) 대한약학회 1996 약학회지 Vol.40 No.4
The anti-Ex-A IgG was specifically labeled with stable isotopes, DL-His-2,4-d2, L-Phe-d5, L-Trp-d5, L-Tyr-2,6-d2 and L-[1-13C]Trp, by growing hybridoma cell in serum-free medium. By use of NMR spectroscopy with selectively labeled Fab fragment, we applied a paratope mapping on antigen-antibody complex. Assignments of the observed carbonyl carbon resonances have been determined by using 13C-15N double labeling method in order to assign the Trp resonances. Photo CIDNP was also applied to investigate the antigen-binding site(s) on the surface residues of antibody. We found that Trp 36, which is located at the VH domain, is an important residue to bind to Ex-A, however, two Tyr on the surface of anti-Ex-A IgG plays no crucial role to bind to antigen. On the basis of these results, we demonstrate that stable isotope-aided NMR strategy can be extended to molecular structural analyses of the complex of an Fab fragment and a protein antigen.
Wheat Germ Agglutinin-conjugated Praecoxin A의 제작 및 특성
김완수(Wan Soo Kim),김만석(Man Suk Kim),김범수(Bum Soo Kim),이민원(Min Won Lee),이도익(Do Ik Lee),최영욱(Yong Wook Choi),김은주(Eun Joo Kim),김하형(Ha Hyung Kim) 대한약학회 2001 약학회지 Vol.45 No.3
Wheat germ agglutinin (WGA) lectin, which binds to human melanoma cell line, was con- jugated with Praecoxin A using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide as a cross-linking agent. Physical mixture(PM) of WGA and Praecoxin A was also prepared by using a non-specific binding property of Praecoxin A to WGA. The WGA : Praecoxin A ratio in the conjugate and PM was approximately 1:18 and 1:20, respectively. The results of hemagglutination assay and enzyme-linked lectin assay indicated that the conjugate and PM maintained the lectin-like properties of the WGA. The binding ratio of coniugate was about 70% during 4-24 hr but the most of Praecoxin A was released within 24 hr in the carte of PM. These results lead to the conclusion that the conjugate is potentially useful for the formulation of injection that requires targeting for melanoma as well as sustained release at the site.
배찬형,김주철,김유정,김상구,나광흠,박병태,김하형,Bae, Chan-Hyung,Kim, Ju-Cheol,Kim, Yu-Jeong,Kim, Sang-Gu,Na, Kwang-Heum,Park, Byung-Tae,Kim, Ha-Hyung 대한약학회 2007 약학회지 Vol.51 No.4
We previously reported the isolation of a sialic acid-specific lectin eluted from the bark of Maackia fauriei using alkaline buffer on a fetuin-affinity column. Application of a borate-based elution buffer in the present study increased the specific activity of purified lectin from crude protein extract by 2.6-fold, whilst only slightly decreasing the recovery by 1.13%. The biological properties of the lectin eluted with borate buffer were the same as those of the lectin eluted with alkaline buffer such as in terms of the hemagglutination activity, hemagglutination inhibition activity, molecular mass, purity, and cytotoxicity to human breast cancer cells. A prepared biotin-labeled lectin conjugate was used to investigate the binding to various glycoproteins. Our results indicate that eluting with borate buffer is more efficient than using alkaline buffer to isolate the lectin adsorbed in a fetuin-affinity column.
김하나(Ha Na Kim),정우혁(Woo Hyuk Chung),배찬영(Chan Hyung Bae),황광우(Kwang Woo Hwang),김하형(HaHyung Kim) 대한약학회 2006 약학회지 Vol.50 No.3
A calcium-dependent sialic acid-binding lectin has been isolated by thyroglobulin-affinity chromatography from the coastal crab Macrophthalmus Japonicus. This lectin, Macrophthalmus Japonicus lectin (MJL), was eluted with 50mM Tris-HCl, 0.3M NaCl, 10mM EDTA, and the recovery yield from the crude protein extract was about 5.6%. The molecular weight of MJL was estimated as 65 kDa in SDS-PAGE both under reducing and non-reducing conditions. MJL induced an agglutination reaction in rabbit, rat, and mouse erythrocytes, but not in human ABO types. This activity was effectively inhibited by sialoglycoproteins such as fetuin, bovine submaxillary mucin, and thyroglobulin.
면역글로부린 G 생성에 대한 아이오타 - 카라기난의 어쥬번트 효과
구본웅,이광재,김만석,김하형,Koo, Bon-Woong,Lee, Kwang-Jae,Kim, Man-Suk,Kim, Ha-Hyung 대한약학회 2000 약학회지 Vol.44 No.6
To investigate the effects of iota-carrageenan (CAR) and/or alum on the adjuvancity as well as the structural difference of oligosaccharide on the IgG2b in the adjuvant effect, C57BL/6 mice were immunized twice with fetuin as a model antigen. CAR alone showed no significant effect on induction of antibody except IgG1. In contrast, Alum-CAR (after mixing of antigen-Alum, CAR adjuvant was prepared) and CAR-Alum (after formulation of antigen-CAR, Alum adjuvant was prepared) enhanced production of antibody, especially, IgG2b. After separation of IgG2b, changes of glycosylation were investigated using enzymelinked lectin assay. High affinity of IgG2b to N-acetylneuraminic acid, galactose and mannose-specific lectin were induced by CAR-Alum adjuvant, however, the affinity of IgG2b induced by CAR-Alum to GlcNAc and GalNAc-specific lectin were much less than that induced by Alum-CAR.
Macrophage의 IL-1β Gene Expression 대한 Lectin-conjugated Ellagitannin의 효과
김한준(Han Jun Kim),김민수(Min Soo Kim),이민원(Min Won Lee),최영욱(Yong Wook Choi),김하형(Ha Hyung Kim),이도익(Do Ik Lee) 대한약학회 2002 약학회지 Vol.46 No.3
Lectin-conjugated praecoxin A, which has developed as a missile antitumor drug, is the one that has conjugated with wheat germ agglutinin (WGA), a kind of carbohydrate- binding protein (lectin) especially bound to melanoma. Praecoxin A is a kind of tannin extracted and purified from plants. Beside this direct antitumor effect as tannins, we have examined an activation of macrophage by lectin-conjugated praecoxin A. We also confirmed the gene expression of IL-1β, both in vitro and in vivo. We added 1,10,100μg/ml of lectin-conjugated praecoxin A, 10μg/ml of pectin,10μg/ml of praecoxin A to normal murine macrophage and analyzed the extracted total RNAs by RT- PCR after 4,8, 12,24 hours. Our data demonstrated that pectin-conjugated praecoxin A increased IL-1β, mRNA expression in a dose dependent manner. However, the effectiveness of lectin-conjugated praecoxln A was not superior to pectin and praecoxin A.
김범수(Bum Soo Kim),이광재(Kwang Jae Lee),이종혁(Jong Hyuk Lee),조두현(Due Hyeon Cho),김윤중(Youn Jung Kim),정병욱(Byung Wook Jung),김하형(Ha Hyung Kim) 大韓藥學會 2002 약학회지 Vol.46 No.1
The present study investigated the rheological properties and adjuvant activity of IgG production of L-Carrageenan raHeenans prepared using two different methods incubation at 37.4 of after either being heated or not being heated at 60o C for 10 minutes. The canageenan exhibited plastic flow with thixotropy in both cases, but the viscosity of the sample that had been heated was significantly higher, C57BL/6 mice were then immunized intraperitoneally with either of the carrageenans containing protein antigens. The maximum adjuvant effect was found when using the nonheated carrageenan, especially in terms of IgG2a, IgG2b, and IgG3 production. In contrast, IgGl antibody production was the same for both types of carrageenan. Furthermore, when administered orally; neither type of carrageenan resulted in the production of any antibodies.
Pedunculagin의 NK cell에 대한 활성화와 흑색종의 전이 억제 효과
이도익(Do Ik Lee),김형근(Hyung Keun Kim),이민원(Min Won Lee),최영욱(Young Wook Choi),김하형(Ha Hyung Kim),김은주(Eun Joo Kim) 대한약학회 2000 약학회지 Vol.44 No.2
Pedunculagin is an ellagitannin purified from Alnus hirsuta var. microphylla, Betulaceae. The effects of pedunculagin on immune system have been characterized to induce enhancement of NK (natural killer) cell cytotoxicities against tumor cells. Here, we report the evaluation of the effects of pedunculagin on the growth of murine B16-FlO melanoma in vivo. After the intradermal inoculation of B16-FlO melanoma, B16-FlO tumors grew progressively in immunocompetent syngenic C57BL/6 mice. The mice treated with pedunculagin(10mg/kg, every 48hrs) resulted in a significant improvement in survival. Inhibitory effects of pedunculagin on lung metastasis in C57BL/6 mice were also detected. Summarizing treatment with pedunculagin has a significant antitumor effect upon B16-FlO murine melanoma.
Maackia fauriei 유래 렉틴의 중성당 및 아미노당 조성
나광흠,박병태,박재완,한경진,박현주,김하형,Na, Kwang-Heum,Park, Byung-Tae,Park, Jae-Wan,Han, Kyong-Jin,Park, Hyun-Joo,Kim, Ha-Hyung 대한약학회 2009 약학회지 Vol.53 No.1
The glycosylation of therapeutic glycoproteins can affect their efficacy, stability, solubility, and half-life. Analyzing the composition of monosaccharides, such as that of neutral and amino sugars, is the first step for elucidating the structure of glycan attached to glycoproteins. In the present study, neutral and amino sugars of lectin obtained from Maackia fauriei were analyzed using an enzyme-linked lectinsorbent assay (ELLA) and high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD). Peroxidase-labeled lectins such as concanavalin A, Ricinus communis agglutinin, and soybean agglutinin were used for ELLA, since they specifically bind to the monosaccharide residue most frequently encountered in a glycan. The hydrosylate of lectin was prepared by treatment with trifluoroacetic acid, which resulted in the lectin mainly possessing the N-glycan consisting of 98.1 pmol Fuc, 342.1 pmol GlcN, 51.9 pmol Gal, 678.9 pmol Man, and 330.7 pmol Xyl. The present results demonstrate that ELLA and HPAEC-PAD are very effective methods for rapidly estimating the types and relative amounts of monosaccharides in intact glycoproteins.