• Title/Summary/Keyword: Transferrin receptor

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Transport of Transferrin-Horseradish Peroxidase Conjugate Through Cultured Caco-2 Cell Monolayer (배양 Caco-2 세포 단층막 실험계에서 트란스페린과 옥시다아제효소 포합체의 세포막투과)

  • Kim, Dong-Chool;Kim, Jie-Hae
    • Journal of Pharmaceutical Investigation
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    • v.29 no.4
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    • pp.287-293
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    • 1999
  • Transport study of horseradish peroxidase and transferrin-horseradish peroxidase conjugate was performed using an in vitro Caco-2 cell cultured monolayer grown on a polycarbonate membrane of $Transwell^{\circledR}$, Horseradish peroxidase was not transported across Caco-2 cell monolayer. Transferrin-horseradish peroxidase conjugate was transported through Caco-2 cell monolayer. The apparent membrane permeability coefficient $(P_{app})$ of transferrin horseradish peroxidase conjugate was $6.54{\times}10^{-7}\;cm/sec$. The $P_{app}$ value of transferrin-horseradish peroxidase conjugate across Caco-2 cell monolayer was increased to $11.9{\times}10^{-7}\;cm/sec$ in the presence of $50\;{mu}g/ml$ brefeldin-A. These results suggest the transferrin receptor mediated transcytosis of transferrin-horseradish peroxidase conjugate across Caco-2 cell monolayer.

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The Determination of Blood-Brain Barrier Permeability and Pharmacokinetics of a Rat Transferrin Receptor Monoclonal Antibody by Brain Perfusion Method and Intravenous Injection Technique in Mice (마우스에서 뇌관류법과 정맥투여법에 의하여 흰쥐 트란스페린 단일항체의 체내동태 및 혈액-뇌 관문 투과성의 검토)

  • 강영숙
    • Biomolecules & Therapeutics
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    • v.10 no.1
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    • pp.37-42
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    • 2002
  • Brain drug targeting through the blood-brain barrier (BBB) in vivo is possible with peptidornirnetic monoclonal antibodies that undergo receptor-mediated transcytosis through the BBB. Monoclonal antibody to the rat transferrin receptor, such as the OX26 was studied in rats as a transport vector through BBB on the transferrin receptor. But, OX26 is not an effective brain delivery vector in mouse. In the present studies, rat monoclonal antibody, 8D3 to the mouse transferrin receptor were evaluated for brain drug targeting vector intransgenic mouse model. Pharrnacokinetic parameters in plasma and organ uptakes were determined at varioustimes after i.v. bolus injection of [$^{}125}I$] 8D3 in Balb/c mice. Brain uptake of [$^{}125}I$] 8D3 was also studied with an internal carotid artery perfusioncapillary depletion method. After i.v. injection of [$^{}125}I$] 8D3, plasma concentrations declined biexponentially with elimination half lift of approximately 2.2 hours. Brain uptake of [$^{}125}I$] 8D3 was $0.50{\pm}0.09$ persent of injected dose per g brain after 2 hours i.v. injection. After perfusion 5 min the apparent volume of distibution of [$^{}125}I$] 8D3 in brain was $22.3 {\mu}l/g,$ which was 4.8 fold higher than the intravascular volume. These studies indicate rat monoclonal antibody to the mouse transferrin receptor, 8D3 may be used for brain drug targeting vector in mice.

Identification of a Transferrin Receptor-binding Peptide from a Phage-displayed Peptide Library (파지-펩타이드 문고로부터 트랜스페린 수용체에 결합하는 펩타이드 탐색)

  • Kim, Sung-Il;Choi, Suk-Jung
    • Journal of Life Science
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    • v.18 no.3
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    • pp.298-303
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    • 2008
  • Using a phage peptide library approach, we have isolated a peptide ligand that binds to transferrin receptor on the surface of human melanoma cell, B16F10. The library was first screened twice by recovering internalized phages and was further screened three times by competitively eluting transferrin receptor-specific phages with human transferrin among the phages bound to the cell surface. The peptides displayed by the selected phages were fused to translocation and catalytic domain of Pseudomonas exotoxin to prepare recombinant toxins. After estimating cytotoxicity of each recombinant toxin toward B16F10 cell, seven clones were selected. Sequence analysis revealed that one of the clones displayed a peptide which had a significant sequence homology with human transferrin. The peptide was chemically synthesized and was shown to be functional in delivering cytotoxic agents into B16F10 cell via interaction with transferrin receptor.

Alteration in the Transferrin Receptor during the Chick Myoblast Fusion in Culture (계배 근원세포의 융합에 따른 Transferrin 수용체의 변화)

  • 이창호;유병제;전영주;정진하;하두봉
    • The Korean Journal of Zoology
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    • v.32 no.2
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    • pp.163-175
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    • 1989
  • Transferrin (Tf) has been known to exert profound effect on the myoblast differentiation in uirto. Therefore, the changes in the amount and affinity of the Tf receptor would accompany the myoblast differentiation. To investigate this possibility, we exarnined the afteration pattern in the level of the Tf receptor during the myoblast fusion. The level of Tf receptor was assayed by measuring the bound 125 I-Tf onto the surface of cultured myoblasts, and it was known that the level of Tf receptor reached the maximum at about 12 hr before the initi ation of the myoblast fusion and decreased as the differentiation proceeded, and that the affinity of Tf receptor to Tf was also decreased. In addition, various inhibitiors of the myoblast fusion also influenced the level of the Tf receptor. Accroding to these results, it is postulated that the level of Tf receptor is highly regulated during the myoblast differentiation.

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Growth Factor Receptor Expression on Brain Tumor Cell Lines : Preliminary Study for in vitro and in vivo Experiments of Immunotoxin Therapy (뇌종양세포주에서의 성장인자수용체의 발현 : 면역독소 치료의 연구를 위한 예비실험)

  • im, Ki-Uk;Ni, Hsiao-Tzu;Low, Walter C.;Hall, Walter A.
    • Journal of Korean Neurosurgical Society
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    • v.29 no.6
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    • pp.731-737
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    • 2000
  • Objective : Growth factor receptors on the tumor cells are known to be expressed highly allowing the tumor cells to bind growth factors to stimulate cellular division. Immunotoxin therapy is one of the novel approaches to the primary malignant brain tumor, and expression of cell-surface receptor is essential for the immunotoxin to have specific anti-tumor activity. Despite promising cytotoxic activity of immunotoxin, tumor responses are not curative on clinical trials, and additional studies are needed regarding various factors influencing the efficacy of the immunotoxin. The purpose of this study is to detect the expression of various growth factor receptors on brain tumor cell lines which are going to be used in these studies. Materials and Methods : The authors detected transferrin receptor(TR), insulin-like growth factor-1 receptor(IGF-1R), and interleukin-4 receptor(IL-4R) on medulloblastoma cell line(Daoy) and glioblastoma cell lines(U373 MG and T98 G) by flow cytometric analysis. Results : TR was expressed on Daoy, U373 MG, and T98 G. IGF-1R was expressed on Daoy and U373 MG, but not on T98 G. IL-4R was expressed on all cell lines tested. Conclusion : The transferrin and interleukin-4 receptors might be good targets for immunotoxin therapy. The results should be considered in additional in vitro and in vivo studies regarding immunotoxin and in establishing the proper treatment model of the immunotoxin therapy including selection of the adequate immunotoxin.

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Cloning and Characterization of the Putative Transferrin Receptor cDNA from the Olive Flounder (Paralichthys olivaceus)

  • Won Kyoung-Mi;Park Soo-Il
    • Fisheries and Aquatic Sciences
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    • v.6 no.2
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    • pp.101-104
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    • 2003
  • A cDNA clone for the olive flounder (Paralichthys olivaceus) transferrin receptor (fTfR) was isolated from a leukocytes cDNA library. The fTfR gene consisted of 2,319 bp encoding 773 amino acid residues. The amino acid sequence alignment of the fTfR showed that their size and hydrophobic profile are similar. In addition, the Tyr-Thr-Arg-Phe (YTRF) motif that is the recognition signal for high-efficiency endocytosis, is conserved very well. This motif is important for functional properties of TfR. The deduced amino acid sequence had $42.4-42.9\%$ identities with the previously reported TfRs of vertebrates. The fTfR was expressed in the blood, kidney, spleen, and liver of healthy olive flounder by the Northern blot hybridization.

Changes in the Level of Transferrin Receptor during the Differentiation of Chick Brain Neuroblasts (계배 신경아세포의 분화에 따른 Transferrin 수용체의 변화)

  • 이창호;전영주
    • The Korean Journal of Zoology
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    • v.35 no.2
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    • pp.144-148
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    • 1992
  • Insulin and transfenin (Tf) were found to be essential for suwival and differentiation of brain neuroblasts obtained from chick embryo. This requirement, however, is Changed from insulin to Tf upon neuronal development of the embryo, and this phenomenon is due to the changes in the levels of corresponding receptors. Using cultured neuroblasts, the level of Tf receptor is also found to increase Lvhile that of insulin receptor falls dramatically during the course of the cell differentiation. These results suggest that the development-specific changes in the levels of insulin and Tf receptors in embryo can be reproduced in the culture system during the differentiation period. Because the culture system used was a defined medium and contained no other macromolecules than insulin and Tf, it appears possible that insulin and Tf may act as signalling molecules in the control of neuronal development.

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The usefulness of soluble transferrin receptor in the diagnosis and treatment of iron deficiency anemia in children

  • Yoon, Se Hoon;Kim, Dong Sup;Yu, Seung Taek;Shin, Sae Ron;Choi, Du Young
    • Clinical and Experimental Pediatrics
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    • v.58 no.1
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    • pp.15-19
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    • 2015
  • Purpose: Soluble transferrin receptor (sTfR) is a truncated extracellular form of the membrane transferrin receptor produced by proteolysis. Concentrations of serum sTfR are related to iron status and erythropoiesis in the body. We investigated whether serum sTfR levels can aid in diagnosis and treatment of iron deficiency anemia (IDA) in children. Methods: Ninety-eight patients with IDA were enrolled and were classified according to age at diagnosis. Group 1 comprised 78 children, aged 6-59 months, and group 2 comprised 20 adolescents, aged 12-16 years. Results: In group 1, patients' serum sTfR levels correlated negatively with mean corpuscular volume; hemoglobin (Hb), ferritin, and serum iron levels; and transferrin saturation and positively with total iron binding capacity (TIBC) and red cell distribution width. In group 2, patients' serum sTfR levels did not correlate with ferritin levels and TIBC, but had a significant relationship with other iron indices. Hb and serum sTfR levels had a significant inverse relationship in both groups; however, in group 1, there was no correlation between Hb and serum ferritin levels. In 30 patients of group 1, serum sTfR levels were significantly decreased with an increase in Hb levels after iron supplementation for 1 month. Conclusion: Serum sTfR levels significantly correlated with other diagnostic iron parameters of IDA and inversely correlated with an increase in Hb levels following iron supplementation. Therefore, serum sTfR levels can be a useful marker for the diagnosis and treatment of IDA in children.

Novel Trimeric Complex for Efficient Uptake of Plasmid Vector into HepG2 Cells

  • Joo, Jong-Hyuck;Park, Jong-Gu
    • Biomedical Science Letters
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    • v.9 no.2
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    • pp.67-74
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    • 2003
  • Viral and non-viral vectors have been used in the delivery of genetic materials into animal cells and tissues, with each approach having pros and cons. Non-viral vectors have many useful merits such as easy preparation, low immunity and size tolerance of a transgene when compared to those of viral vectors. Delivery specificity may be achieved by complex formation between receptor ligands and a non-viral vector. In the present study, non-viral vector systems are investigated in an effort to find a practical delivery means for gene therapy, Receptor-ligand interaction between transferrin-receptor and transferrin was utilized for efficient gene transfer into cancer cells. A plasmid vector, pcDNA3 (LacZ) was ligated with a small duplexed oligo fragment in which a Biotin- VN$^{TM}$ phosphoramidite was placed in the middle of the oligo. The plasmid vector labeled by biotin was then conjugated with biotin-labeled transferrin via streptavidin. This trimeric conjugates were delivered to a hepatoma cell line, HepG2. The delivery efficiency of the trimeric conjugate was 2-fold higher than that of cationic liposomes used for transfection of a plasmid vector. These results demonstrate that a plasmid vector can be efficiently transferred into cells by forming a trimeric complex of plasmid vector-linker-ligand.

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Immunoradiometric Assay using Monoclonal Antibody Against Human Serum Transferrin Receptor for Diagnosis of Iron Deficiency (사람 혈청 트란스페린수용체의 단클론 항체를 이용한 방사면역측정과 철영양상태의 진단)

  • 김승렬
    • Journal of Nutrition and Health
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    • v.29 no.9
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    • pp.971-980
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    • 1996
  • The soluble transferrin receptor(TfR) in human serum has been shown recently to be a truncated form of intact membrane bound receptor containing most of the extracellular domain. We purfied the transferin-free TfR from human serum by immounoaffinity chromatography which produced the single protein identity in high resolution gel chormatography. The monoclonal antibodies(MAb) against purifed serum TfR were produced by fusion of spleen cells o fimmunized Balb/c mice and SP2 cells. Ten hybrids producing MAb specific for serum TfR were identifed and determine their iostypes. A immunoraddiometric assay (IRMA) for serum TfR was established using two monoclonal IgG1 antibodies as the coating and indicator antibodies on the bosis of their suitability in sandwich IRMA of serum TfR. The mean serum TfR levels in the 15 normal male, 15 normal female, and 19 iron-deficient subjects were 5.4$\pm$0.98, 4.6$\pm$0/76, and 18.0$\pm$12.8mg/1, respectively, and the difference in mean values between normal and iron deficient subjects was significant(p=0.0005). There existed the inverse logarithmic relationship(r=-0.9336, p<0.0001) between the serum TfR and ferritin levels.

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