• Title/Summary/Keyword: single chain

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Function of the Tethered rec-eCG in Rat and Equine Receptors

  • Park, Jong-Ju;Jargal, Naidansuren;Yoon, Jong-Taek;Min, Kwan-Sik
    • Reproductive and Developmental Biology
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    • v.33 no.4
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    • pp.229-236
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    • 2009
  • The glycoprotein hormone family represents a class of heterodimers, that includes the placental hormone equine chorionic gonadotropin (eCG) and the anterior pituitary hormones- follitropin (FSH), lutropin (LH), and thyrotropin (TSH). The 4 hormones are heterodimers, with a common $\alpha$-subunit and unique $\beta$-subunits. eCG is the most heavily glycosylated of the known pituitary and placental glycoprotein hormones. Recent observations using single chain glycoprotein hormone analogs in which, the $\beta$-and $\alpha$-subunits are linked, implied that heterodimeric-like quaternary configuration is not a prerequisite for receptor/signal transduction. To study the function and signal transduction of tethered rec-eCG, a single chain eCG molecule was constructed and rec-eCG protein was produced. Molecular mass of the single chain is about 45 kDa. All mice were ovulated by tethered rec-eCG treatment. The dual activity of tethered rec-eCG was determined in receptor cell lines of nonequid species; in fact, this dual activity was proven in species other than horse. Tethered rec-eCG in equids does not bind to FSH receptors, suggesting that eCG is primarily an LH-like hormone in the horse. Taken together, these data suggest that tethered rec-eCG has dual activity in nonequid species in vitro. However, it has only LH-like activity in equid species in vitro.

Performance Analysis of Opportunistic Spectrum Access Protocol for Multi-Channel Cognitive Radio Networks

  • Kim, Kyung Jae;Kwak, Kyung Sup;Choi, Bong Dae
    • Journal of Communications and Networks
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    • v.15 no.1
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    • pp.77-86
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    • 2013
  • Cognitive radio (CR) has emerged as one of effective methods to enhance the utilization of existing radio spectrum. Main principle of CR is that secondary users (SUs) are allowed to use the spectrum unused by primary users (PUs) without interfering PU's transmissions. In this paper, PUs operate on a slot-by-slot basis and SUs try to exploit the slots unused by PUs. We propose OSA protocols in the single channel and we propose an opportunistic spectrum access (OSA) protocols in the multi-channel cognitive radio networks with one control channel and several licensed channels where a slot is divided into contention phase and transmission phase. A slot is divided into reporting phase, contention phase and transmission phase. The reporting phase plays a role of finding idle channels unused by PUs and the contention phase plays a role of selecting a SU who will send packets in the data transmission phase. One SU is selected by carrier sense multiple access / collision avoidance (CSMA/CA) with request to send / clear to send (RTS/CTS) mechanism on control channel and the SU is allowed to occupy all remaining part of all idle channels during the current slot. For mathematical analysis, first we deal with the single-channel case and we model the proposed OSA media access control (MAC) protocol by three-dimensional discrete time Markov chain (DTMC) whose one-step transition probability matrix has a special structure so as to apply the censored Markov chain method to obtain the steady state distribution.We obtain the throughput and the distribution of access delay. Next we deal with the multi-channel case and obtain the throughput and the distribution of access delay by using results of single-channel case. In numerical results, our mathematical analysis is verified by simulations and we give numerical results on throughput and access delay of the proposed MAC protocol. Finally, we find the maximum allowable number of SUs satisfying the requirements on throughput and access delay.

Live Cell Detection of Monoclonal Antibody Light and Heavy Chain mRNAs using Molecular Beacons (분자 비컨을 이용한 살아 있는 세포에서 단일클론항체 경쇄와 중쇄 mRNA 검출에 의한 세포주 선별방법)

  • Jeong, Seunga;Rhee, Won Jong
    • KSBB Journal
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    • v.31 no.1
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    • pp.33-39
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    • 2016
  • Developing the method for the selection of animal cell line producing therapeutic monoclonal antibody (mAb) is invaluable as its market is rapidly growing. Although the quality of produced mAb is as important as quantity, however there is no method developed for the selective screening of cell lines on the basis of both quantity and quality. From recent reports, the ratio of light and heavy chain mRNAs of mAb in the cell is a key parameter for the indication of product quality. Therefore, it is obvious that developing the novel method that can detect both light and heavy chain mRNAs in single live cell will provide unprecedented opportunities in bio-industry. Here, we have constructed oligonucleotide probes, molecular beacons for the detection of light or heavy chain mRNAs, respectively, in the live cells producing mAbs. Both beacons showed increased fluorescent intensity after transient transfection of plasmid expressing mAbs analyzed by fluorometer. Flow cytometric analysis clearly demonstrated that both molecular beacons can simultaneously detect the expression of light and heavy chain mRNAs of mAb in the same cell. The technique described in the thesis provides the new direction and concept for developing the method for the smart selection of cell lines producing recombinant proteins including therapeutic mAbs.

Optimal Operation for Green Supply Chain Considering Demand Information, Collection Incentive and Quality of Recycling Parts

  • Watanabe, Takeshi;Kusukawa, Etsuko
    • Industrial Engineering and Management Systems
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    • v.13 no.2
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    • pp.129-147
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    • 2014
  • This study proposes an optimal operational policy for a green supply chain (GSC) where a retailer pays an incentive for collection of used products from customers and determines the optimal order quantity of a single product under uncertainty in product demand. A manufacturer produces the optimal order quantity of product using recyclable parts with acceptable quality levels and covers a part of the retailer's incentive from the recycled parts. Here, two scenarios for the product demand are assumed as: the distribution of product demand is known, and only both mean and variance are known. This paper develops mathematical models to find how order quantity, collection incentive of used products and lower limit of quality level for recycling affect the expected profits of each member and the whole supply chain under both a decentralized GSC (DGSC) and an integrated GSC (IGSC). The analysis numerically compares the results under DGSC with those under IGSC for each scenario of product demand. Also, the effect of the quality of the recyclable parts on the optimal decisions is shown. Moreover, supply chain coordination to shift the optimal decisions of IGSC is discussed based on: I) profit ratio, II) Nash bargaining solution, and III) Combination of (I) and (II).

Kinetics of Converting Single Chain Urokinase Type Plasminogen Activator into Two Chain Plasminogen Activator in Cultivating HEK Cells with Low Serum Containing Medium (저혈청 배지에서 생산된 scu-PA의 tc-PA로의 전환에 관한 연구)

  • 김영남;김현구
    • KSBB Journal
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    • v.9 no.1
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    • pp.48-54
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    • 1994
  • A modified amidolytic assay and a fibrin plate method were used to accurately measure the concentration of single chain urokinase type plasminogen activator (scu-PA) and two-chain urokinase type plasminogen activator (tc-PA) in the spent media. $1.65{\times}10^6$(viable cells/ml) of maximum cell density and 1670(IU/ml) of scu-PA concentration were obtained in 1% serum containing medium. The overall conversion ratio from scu-PA to tc-PA was less than 10%. In the results of batch cultivation in a spinner vessel, $4.43{\times}10^6(total cells/ml)$ of maximum cell density and 1560(IU/ml) of scu-PA concentration was observed. The maximum scu-PA concentration and specific scu-PA Productivity were obtained in 1760(IU/ml) and $3.13{\times}10^{-4}(IU/cell)$, respectively, from perfusion cultivation. The conveysion ratios from batch, fed-batch and perfusion cultivations were less than 12%, which means that about 90% of scu-PA secreted from the cells can be maintained during the cultivations.

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Single-tube nested reverse transcription-polymerase chain reaction for simultaneous detection of genotyping of porcine reproductive and respiratory syndrome virus without DNA carryover contamination (DNA 교차오염 방지기능이 있는 single-tube nested reverse transcription-polymerase chain reaction을 이용한 돼지생식기호흡기증후군바이러스 유전형 감별진단)

  • Jeong, Pil-Soo;Park, Su-Jin;Kim, Eun-Mi;Park, Ji-Young;Park, Yu-Ri;Kang, Dae-Young;Cha, Hyun-Ouk;Lee, Kyoung-Ki;Kim, Seong-Hee;Park, Choi-Kyu
    • Korean Journal of Veterinary Service
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    • v.39 no.2
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    • pp.107-116
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    • 2016
  • In the study, we developed and evaluated a uracil N-glycosylase (UNG)-supplemented single-tube nested reverse transcription-polymerase chain reaction (UsnRT-PCR) assay that can carried out first-round RT-PCR and second-round nested PCR in a reaction tube without reaction tube opening and can simultaneously detect EU- and NA-PRRSV. The UsnRT-PCR confirmed to have a preventing ability of mis-amplification by contamination of pre-amplified PRRSV DNA from previous UsnRT-PCR. Primer specificities were evaluated with RNAs extracted from 8 viral strains and our results revealed that the primers had a high specificity for both genotypes of PRRSV. The sensitivity of the UsnRT-PCR was 0.1 $TCID_{50}$/0.1 mL for EU- or NA-PRRSV, respectively, which is comparable to that of previously reported real time RT-PCR (RRT-PCR). Clinical evaluation on 110 field samples (60 sera and 50 lung tissues) by the UsnRT-PCR and the RRT-PCR showed that detection rates of the UsnRT-PCR was 70% (77/110), and was relatively higher than that of the RRT-PCR (69.1%, 76/110). The percent positive or negative agreement of the UsnRT-PCR compared to RRT-PCR was 96.1% (73/76) or 90.9% (30/33), showing that the test results of both assays may be different for some clinical samples. Therefore, it is recommend that diagnostic laboratory workers use the two diagnostic assays for the correct diagnosis for the relevant samples in the swine disease diagnostic laboratories. In conclusion, the UsnRT-PCR assay can be applied for the rapid, and reliable diagnosis of PRRSV without concerns about preamplified DNA carryover contamination that can occurred in PCR process in the swine disease diagnostic laboratories.

The Improved Antigen-binding Activity of Biosimilar Remicade ScFv Antibodies by Fusion of the Leucine Zipper Domain (Leucine zipper도메인의 융합에 의한 바이오시밀러 레미케이드 Single-chain Fv 항체의 항원 결합력 개선)

  • Kim, Jin-Kyoo;Kim, Tae Hwan
    • Journal of Life Science
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    • v.30 no.11
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    • pp.1012-1020
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    • 2020
  • Remicade is a therapeutic biosimilar natural antibody in which the mouse variable domain has been linked to the human constant domain. It is a chimeric monoclonal antibody specific to tumor necrosis factor-alpha (TNF-α) and has been developed for the treatment of rheumatoid arthritis. To investigate the biological activity of the Remicade antibody, we carried out a bioinformatics study using a protein data bank to characterize the TNF-α antigen binding mechanism of the Remicade natural antibody. Because the production of the Remicade antibody is often limited by genetic instability of the natural antibody-producing cell, we generated a Remicade single-chain variable domain fragment antibody (Remicade) in which a heavy chain variable domain (VH) is joined with a light chain variable domain (VL) by a polypeptide linker. Furthermore, Remicade was fused to a leucine zipper (RemicadeScZip) for higher production and higher antigen-binding activity than Remicade. The Remicade and Remicade ScZip were expressed in Escherichia coli and purified by a Ni+-NTA-agarose column. As expected, the purified proteins had migrated as 28.80 kDa and 33.96 kDa in sodium dodecyl sulfate-polyacrylamide electrophoresis. The TNF-α antigen binding activity of Remicade was not observed by ELISA and western blot. In contrast, RemicadeScZip showed antigen-binding activity. Additional bio-layer interferometry analysis confirmed the antigen-binding activity of RemicadeScZip, suggesting that the leucine zipper stabilized the folding of RemicadeScZip in a denatured condition and improved the TNF-α antigenbinding activity.

In vitro Propagation of Transgenic Ginsengs Introduced with Ferritin Light Heavy Chain Gene through Single Embryo Culture (Ferritin Light Heavy Chain 유전자가 도입된 인삼형질전환체의 단일배발생을 통한 식물체의 기내증식)

  • 윤영상;김종학;김무성;양덕춘
    • Korean Journal of Plant Resources
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    • v.17 no.2
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    • pp.161-168
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    • 2004
  • Optimal regeneration conditions of ginseng transformants were studied. It has been known that Ferritin Light Heavy Chain (FLHC) gene remove the several heavy metal by combination, store and transport. To obtain the ginseng tolerant to heavy metal, binary vector was introduced in Agrobacterium by tri-parental mating and then Agrobacterium tumefaciens MP90/FLHC was selected on the AB media and MS media containing kanamycin. Explants were co-cultured with Agrobacterium tumefaciens MP90/FLHC, which contained NPT II as a selectable marker, tadpole ferritin heavy chain (FLHC) gene and human ferritin light chain gene and then a number of embryos were induced. The induced embryo transferred to shooting media consisting of MS medium supplemented with GA 10 mg/L. As a result of examination that induced the normal growth of transfomants, transformants showed the equivalent growth in both root and shoot on the media containing the 1/3 MS.

A Scheduling Problem for Production-Delivery in a Supply Chain (생산 및 배송을 위한 공급사슬망에서의 일정계획에 관한 연구)

  • Yoon, Sang-Hum;Lee, Ik-Sun
    • Journal of the Korean Operations Research and Management Science Society
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    • v.31 no.4
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    • pp.35-47
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    • 2006
  • This paper considers an integrated scheduling problem for consecutive production and delivery stages in a two-stage supply chain. The Production is performed on a single facility and then the finished products are delivered to the customer by capacitated multiple vehicles. The objective of this paper is to obtain job sequencing and delivery batching minimizing the total cost of the associated WIP inventory, finished product inventory and delivery. The inventory cost is characterized by the sum of weighted flowtime. The delivery cost is proportional to the required number of delivery batches. Some polynomial-solvable cases are derived. For the general case, two efficient heuristic algorithms are suggested, and then the heuristics are tested through some numerical experiments.

Demand Variability Impact on the Replenishment Policy in a Two-Echelon Supply Chain Model (두 계층 공급사슬 모형에서 발주정책에 대한 수요 변동성 영향)

  • Kim Eungab
    • Journal of the Korean Operations Research and Management Science Society
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    • v.29 no.3
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    • pp.111-127
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    • 2004
  • We consider a supply chain model with a make-to-order production facility and a single supplier. The model we treat here is a special case of a two-echelon inventory model. Unlike classical two-echelon systems, the demand process at the supplier is affected by production process at the production facility as well as customer order arrival process. In this paper, we address that how the demand variability impacts on the optimal replenishment policy. To this end, we incorporate Erlang and phase-type demand distributions into the model. Formulating the model as a Markov decision problem, we investigate the structure of the optimal replenishment policy. We also implement a sensitivity analysis on the optimal policy and establish its monotonicity with respect to system cost parameters.