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Ko, Sanghwan,Hwang, Bora,Na, Jung-Hyun,Lee, Jisun,Jung, Sang Taek American Chemical Society 2019 Journal of agricultural and food chemistry Vol.67 No.43
<P>Despite remarkable contribution of green fluorescent protein and its variants for better understanding of various biological functions, its application for anaerobic microorganisms has been limited because molecular oxygen is essential for chromophore formation. To overcome the limitation, we engineered a plant-derived light, oxygen, or voltage (LOV) domain containing flavin mononucleotide for enhanced spectral properties. The resulting LOV variants exhibited improved fluorescence intensity (20 and 70% higher for SH3 and 70% for BR1, respectively) compared to iLOV, an LOV variant isolated in a previous study, and the quantum yields of the LOV variants (0.40 for SH3 and 0.45 for BR1) were also improved relative to that of iLOV (<I>Q</I> = 0.37). In addition to fluorescence intensity, the identified mutations of SH3 enabled an improved thermostability of the protein. The engineered LOV variants with enhanced spectral properties could provide a valuable tool for fluorescent molecular probes under anaerobic conditions.</P> [FIG OMISSION]</BR>
Ko Sanghwan,Park Sora,Sohn Myung Ho,조미경,Ko Byoung Joon,Na Jung-Hyun,Yoo Hojin,Jeong Ae Lee,Ha Kyungsoo,Woo Ju Rang,Lim Chungsu,Shin Jung Hyu,Lee Dohyun,Choi So-Young,Jung Sang Taek 생화학분자생물학회 2022 Experimental and molecular medicine Vol.54 No.-
The pH-selective interaction between the immunoglobulin G (IgG) fragment crystallizable region (Fc region) and the neonatal Fc receptor (FcRn) is critical for prolonging the circulating half-lives of IgG molecules through intracellular trafficking and recycling. By using directed evolution, we successfully identified Fc mutations that improve the pH-dependent binding of human FcRn and prolong the serum persistence of a model IgG antibody and an Fc-fusion protein. Strikingly, trastuzumab-PFc29 and aflibercept-PFc29, a model therapeutic IgG antibody and an Fc-fusion protein, respectively, when combined with our engineered Fc (Q311R/M428L), both exhibited significantly higher serum half-lives in human FcRn transgenic mice than their counterparts with wild-type Fc. Moreover, in a cynomolgus monkey model, trastuzumab-PFc29 displayed a superior pharmacokinetic profile to that of both trastuzumab-YTE and trastuzumab-LS, which contain the well-validated serum half-life extension Fcs YTE (M252Y/S254T/T256E) and LS (M428L/N434S), respectively. Furthermore, the introduction of two identified mutations of PFc29 (Q311R/M428L) into the model antibodies enhanced both complement-dependent cytotoxicity and antibody-dependent cell-mediated cytotoxicity activity, which are triggered by the association between IgG Fc and Fc binding ligands and are critical for clearing cancer cells. In addition, the effector functions could be turned off by combining the two mutations of PFc29 with effector function-silencing mutations, but the antibodies maintained their excellent pH-dependent human FcRn binding profile. We expect our Fc variants to be an excellent tool for enhancing the pharmacokinetic profiles and potencies of various therapeutic antibodies and Fc-fusion proteins.
An Approach for Synthesis and Characterization of Zirconia Nanopowder via Microwave Thermal Process
Athar, Taimur,Han, Sanghwan,Ko, Taegyung,Lee, Ik-Mo Taylor Francis 2009 Synthesis and reactivity in inorganic, metal-organ Vol.39 No.5
<P> We applied a simple, versatile and an efficient non-toxic methodology based on microwave irradiation process to prepare a gel from n-zirconium butoxide without using any solvent at ambient temperature to open new possibilities to control particle size distribution, surface chemistry and agglomeration. The alkoxy-derived gel changed to tetragonal zirconia powder on heating at 400°C with crystalline particles size (3 ∼ 5 nm). Zirconia exhibited strong luminescence under a UV laser. The presence of a organic moieties as impurities exhibit a strong luminescence in zirconia powder. From the structural analysis it is concluded that the morphology of the oxide strongly depends on the nature of the precursors and the synthetic methology.</P>
An Integrated Low Temperature Approach for the Synthesis of Titania via Sol-Gel process
Athar, Taimur,Han, Kyusuk,Han, Sanghwan,Ko, Taegyung,Lee, Ik-Mo Taylor Francis 2009 Synthesis and reactivity in inorganic, metal-organ Vol.39 No.5
<P> Colloidal titania particles with nanosize size distribution were prepared via sol-gel process by using oxo-titanium alkoxide as a molecular precursor. The transformation from oxo-titanium alkoxide to metal oxide was studied by IR, UV, thermal analysis, X-ray and TEM. TEM and XRD techniques were used to characterize the microstructure and crystallite morphology. The experimental results show that the titania has a good crystallinity with a particle size (5 nm), shape and with favorable surface morphology. These results support that titania has strong implications for the interaction between the oleic acid used as surfactant and Ti nanoparticle to enhance the stability with controlled functional properties.</P>
A human antibody against human endothelin receptor type A that exhibits antitumor potency
주만석,Ahn Hye-Mi,Han Seong-Gu,Ko Sanghwan,Na Jung-Hyun,조미경,임충수,Ko Byoung Joon,Yu Yeon Gyu,이원규,김연재,Jung Sang Taek 생화학분자생물학회 2021 Experimental and molecular medicine Vol.53 No.-
Endothelin receptor A (ET A ), a class A G-protein-coupled receptor (GPCR), is involved in the progression and metastasis of colorectal, breast, lung, ovarian, and prostate cancer. We overexpressed and purified human endothelin receptor type A in Escherichia coli and reconstituted it with lipid and membrane scaffold proteins to prepare an ET A nanodisc as a functional antigen with a structure similar to that of native GPCR. By screening a human naive immune single-chain variable fragment phage library constructed in-house, we successfully isolated a human anti-ET A antibody (AG8) exhibiting high specificity for ET A in the β-arrestin Tango assay and effective inhibitory activity against the ET-1-induced signaling cascade via ET A using either a CHO-K1 cell line stably expressing human ET A or HT-29 colorectal cancer cells, in which AG8 exhibited IC 50 values of 56 and 51 nM, respectively. In addition, AG8 treatment repressed the transcription of inhibin βA and reduced the ET A -induced phosphorylation of protein kinase B and extracellular regulated kinase. Furthermore, tumor growth was effectively inhibited by AG8 in a colorectal cancer mouse xenograft model. The human anti-ET A antibody isolated in this study could be used as a potential therapeutic for cancers, including colorectal cancer.