Characterization of the Glycan Structures of Glycoprotein GA733-Fc Expressed in a Baculovirus-Insect Cell System

Lu Qiao,이경진,고기성 대한화학회 2015 Bulletin of the Korean Chemical Society Vol.36 No.1

Baculovirus-insect cell systems have been used to express functional recombinant biopharmaceutical proteins. Two pFastBac Dual vectors carrying the gene encoding antigen GA733, a cell-surface glycoprotein, fused to the IgG Fc (GA733-Fc) or KDEL (endoplasmic reticulum [ER] retention sequence) (GA733-FcK) genes were constructed to generate baculoviruses expressing the corresponding recombinant proteins in insect cells. The expression of the recombinant GA733-Fc and GA733-FcK proteins and their glycan structure profiles were assessed under various conditions by analyzing the cell line (Sf9 and High Five), the postinfection (PI) time (48, 72, and 96h), and the harvested sample (cell culture media [CM] or cell lysate [CL]). Immunoblotting showed nonidentical expression of both GA733-Fc and GA733-FcK under various conditions. Glycosylation analysis revealed that varying conditions affected the glycan structure profiles. These results suggest that the PI time, subcellular localization, and cell type affect recombinant protein expression as well as glycosylation in the baculovirus-insect cell system.

Investigation and Simulation Study on the Cascading Trip-off Fault of a Large Number of Wind Turbines in China on May 14, 2012

Qiao, Ying,Lu, Zong-Xiang,Lu, Ji,Ruan, Jia-Yang,Wu, Lin-lin The Korean Institute of Electrical Engineers 2015 Journal of Electrical Engineering & Technology Vol.10 No.6

The integration of the large-scale wind power brings great challenge to the stability of the power grid. This paper investigates and studies the fault on May 14, 2012 of the large-scale cascading trip-off of wind turbines in North China. According to the characteristics of the voltage variation, the fault process is divided into three stages: the pre-event stage, the critical stage before cascading, and the cascading stage. The scenes in the fault are reproduced, using the full-size actual power system model. Simulation models of double-fed induction generators (DFIGs) and SVCs including protection settings and controller strategies are carefully chosen to find out the reason of voltage instability in each stage. Some voltage dynamic that have never been observed before in the faults of the same kind are analyzed in detail, and an equivalent voltage sensitive dynamic model of DFIG is proposed for the fast computation. The conclusions about the voltage dynamics are validated by the actual PMU observation evidence.

먹방 인플루언서 콘텐츠 협찬 고지가 기대불일치에 미치는 영향 : 인플루언서 평판의 조절효과를 중심으로

LU QIAO,류은아 한국광고학회 2021 광고학연구 Vol.32 No.6

Function and optimization of colorectal cancer vaccine candidate spGA733-FcK and spGA733-Fc expressed in baculovirus-insect cell system

Lu Qiao,Kyung-Jin Lee,Kisung Ko 한국응용곤충학회 2013 한국응용곤충학회 학술대회논문집 Vol.2013 No.10

The antigen GA733 is a cell-surface highly expressed glycoprotein on most human colorectal carcinomas. GA733 can be characterized as a cancer vaccine. In this study, GA733 was fused to the human immunoglobulin IgG Fc fragment to become recombinant gene GA733-Fc. Based on this, 4 recombinant genes were constructed as follows: GA733-Fc with signal peptide sequence and fusion of ER retention sequence (KDEL) (spGA733-FcK), GA733-Fc with signal sequence (spGA733-Fc), GA733-Fc fused to ER retention sequence (GA733-FcK) without signal peptide and GA733-Fc without signal peptide. Baculovirus-insect cell expression system is widely used for the high level production of recombinant proteins especially for glycoproteins. Constructed 4 recombinant genes were cloned to baculovirus express vectors. DH10Bac E.coli.-mediated transformation was used to generate recombinant bacmid DNA. Recombinant DNA was confirmed by PCR. Insect cell was transfected by bacmid to produce the recombinant baculovirus infects insect cell to produce recombinant protein. Western blot and sandwich ELISA showed the expression of recombinant proteins. Each cell lines (sf9 and HighFive) differed in recombinant proteins production level and protein secretion capability. N-Glycosylation analysis showed the function of signal peptide and ER retention sequence (KDEL). Taken together, baculovirus-insect cell system can be used to express recombinant GA733-Fc glycoproteins.

Function and glycosylation of colorectal cancer vaccine candidate spGA733-FcK and spGA733-Fc expressed in baculovirus-insect cell system

Lu Qiao,Kyung-Jin Lee,Kisung Ko 한국당과학회 2013 한국당과학회 학술대회 Vol.2013 No.1

The antigen GA733 is a cell-surface highly expressed glycoprotein on most human colorectal carcinomas. GA733 can be characterized as a cancer vaccine. In this study, GA733 was fused to the human immunoglobulin IgG Fc fragment to become recombinant gene GA733-Fc. Based on this, 4 recombinant genes were constructed as follows: GA733-Fc with signal peptide sequence and fusion of ER retention sequence (KDEL) (spGA733-FcK), GA733-Fc with signal sequence (spGA733-Fc), GA733-Fc fused to ER retention sequence (GA733-FcK) without signal peptide and GA733-Fc without signal peptide. Baculovirus-insect cell expression system is widely used for the high level production of recombinant proteins especially for glycoproteins. Constructed 4 recombinant genes were cloned to baculovirus express vectors. DH10Bac E.coli.-mediated transformation was used to generate recombinant bacmid DNA. Recombinant DNA was confirmed by PCR. Insect cell was transfected by bacmid to produce the recombinant baculovirus infects insect cell to produce recombinant protein. Western blot and sandwich ELISA showed the expression of recombinant proteins. Each cell lines (sf9 and HighFive) differed in recombinant proteins production level and protein secretion capability. N-Glycosylation analysis showed the function of signal peptide and ER retention sequence (KDEL). Taken together, baculovirus-insect cell system can be used to express recombinant GA733-Fc glycoproteins.

Investigation and Simulation Study on the Cascading Trip-off Fault of a Large Number of Wind Turbines in China on May 14, 2012

Ying Qiao,Zong-Xiang Lu,Ji Lu,Jia-Yang Ruan,Lin-lin Wu 대한전기학회 2015 Journal of Electrical Engineering & Technology Vol.10 No.6

The integration of the large-scale wind power brings great challenge to the stability of the power grid. This paper investigates and studies the fault on May 14, 2012 of the large-scale cascading trip-off of wind turbines in North China. According to the characteristics of the voltage variation, the fault process is divided into three stages: the pre-event stage, the critical stage before cascading, and the cascading stage. The scenes in the fault are reproduced, using the full-size actual power system model. Simulation models of double-fed induction generators (DFIGs) and SVCs including protection settings and controller strategies are carefully chosen to find out the reason of voltage instability in each stage. Some voltage dynamic that have never been observed before in the faults of the same kind are analyzed in detail, and an equivalent voltage sensitive dynamic model of DFIG is proposed for the fast computation. The conclusions about the voltage dynamics are validated by the actual PMU observation evidence.

p13 from group II baculoviruses is a killing-associated gene

( Nan Lu ),( En Qi Du ),( Yang Kun Liu ),( Hong Qiao ),( Lun Guang Yao ),( Zi Shu Pan ),( Song Ya Lu ),( Yi Peng Qi ) 생화학분자생물학회 2012 BMB Reports Vol.45 No.12

p13 gene was first described in Leucania separata multinuclear polyhedrosis virus (Ls-p13) several years ago, but the function of P13 protein has not been experimentally investigated to date. In this article, we indicated that the expression of p13 from Heliothis armigera single nucleocapsid nucleopolyhedrovirus (Ha-p13) was regulated by both early and late promoter. Luciferase assay demonstrated that the activity of Ha-p13 promoter with hr4 enhancer was more than 100 times in heterologous Sf9 cells than that in nature host Hz-AM1 cells. Both Ls-P13 and Ha-P13 are transmembrane proteins. Confocal microscopic analysis showed that both mainly located in the cytoplasm membrane at 48 h. Results of RNA interference indicated that Ha-p13 was a killing-associated gene for host insects H. armigera. The AcMNPV acquired the mentioned killing activity and markedly accelerate the killing rate when expressing Ls-p13. In conclusion, p13 is a killing associated gene in both homologous and heterologous nucleopolyhedrovirus. [BMB Reports 2012; 45(12): 730-735]

Cecropin Suppresses Human Hepatocellular Carcinoma BEL-7402 Cell Growth and Survival in vivo without Side-Toxicity

Jin, Xiao-Bao,Wang, Ying-Jiao,Liang, Lu-Lu,Pu, Qiao-Hong,Shen, Juan,Lu, Xue-Mei,Chu, Fu-Jiang,Zhu, Jia-Yong Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.13

Conventional chemotherapy against hepatocellular carcinoma typically causes various side effects. Our previous study showed that cecropin of Musca domestica can induce apoptosis in human hepatocellular carcinoma BEL-7402 cells in vitro. However, whether cecropin inhibits BEL-7402 cell in vivo and the question of possible side effects remained undentified. The present study confirmed tumor-inhibitory effects of cecropin in vivo, and furthermore strongly suggested that cecropin cytotoxicity in BEL-7402 cells in vivo may be mainly derived from its pro-apoptotic action. Specifically, we found that cecropin exerted no obvious side effects in tumor-bearing mice as it had no significant hematoxicity as well as visceral toxicity. Therefore, cecropin may be a potential candidate for further investigation as an antitumor agent against hepatocellular carcinoma.

Implementation of the strain hardening model into buffer material in high level waste repository

Lu Chen,Lan Qiao,Ming Cui,Qingwen Li 한국지질과학협의회 2017 Geosciences Journal Vol.21 No.2

Engineered Barrier System (EBS) primarily provides a protective cover for the High Level Waste (HLW) canisters. In addition, it functions as an efficient hydraulic and chemical barrier system. For in situ disposal applications, the ideal buffer material is bentonite and it is fabricated into blocks to build EBS. Currently, traditional methods introduce significant errors in constitutive modeling of those modeled blocks. In this paper, the particular failure laws and stress-strain laws of bentonite blocks were furnished from a lot of uniaxial and triaxial tests. Also, it showed that the stress-strain relations given by the traditional constitutive relations did not correspond to the failure forms of bentonite specimens in the experiments. On this basis, the relations between internal friction angle, cohesion and plastic strain were analyzed so that a new strain hardening model was established under the framework of Mohr-Coulomb criterion. In the end, a series of 3D computation of HLW disposal were carried out using the software FLAC3D, where the proposed strain hardening model led to satisfactory prediction of the buffer materials.

Exogenous Spermidine Promoted Ca2+ Absorption in Lettuce Roots and Reduced the Incidence of Tipburn

Qiao Yang,Na Lu,Lei Wang,Xiao-qing Huang,Da-quan Yang,Jin Sun 한국원예학회 2018 원예과학기술지 Vol.36 No.5

A physiological disorder, called tipburn, commonly occurs in lettuce (Lactuca sativa L.), particularly in greenhouse cultivation and in hot seasons, and reduces its marketability. Calcium (Ca) deficiency is the main cause of tipburn injury. In the present study, we investigated the effect of exogenous spermidine (Spd) on the occurrence of tipburn in hydroponically cultivated head lettuce (var. capitata L.) and elucidated the role of Ca ions (Ca2+) in its prevention with respect to absorption and transport. Ca deficiency in the nutrient solution resulted in reduced uptake of Ca2+ in lettuce roots, thereby resulting in Ca deficiency in leaves, leading to leaf senescence and tipburn. An application of exogenous Spd promoted Ca2+ uptake in lettuce roots under Ca deficiency and promoted Ca2+ transportation to leaves, especially to inner leaves, thus delaying leaf senescence and reducing the incidence of tipburn. The results indicated that application of exogenous Spd is an effective method to decrease tipburn in lettuce caused by Ca deficiency.