• Title/Summary/Keyword: and biotin

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Synthesis and Characterization of a Receptor-Targeting Contrast Agent

  • Yang, Taegyun;Park, Ji-Hyung;Lee, Seung-Cheol;Kim, Cheol-Su;Cho, Jee-Hyun;Lee, Chul-Hyun;Cheong, Chae-Joon
    • Journal of the Korean Magnetic Resonance Society
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    • v.7 no.1
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    • pp.46-54
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    • 2003
  • We synthesized a contrast agent for MRI that is capable of binding to the ABP-1 receptor and enhancing the contrast of the targeted cells. We used a lysine dendrimer (G=3)DTPA[Gd] as the contrast agent and synthesized a biotinylated polyclonal antibody for ABP-1 as the first antibody. Lysine dendrimers were prepared using the solid phase peptide synthesis method.$^3$ Amino-terminated lysine dendrimers were then coupled to DTPA using the anhydride method. Gd was complexed with the DTPA-lysine dendrimer in an acidic solution of 3 eq GdCl$_3$ to one of DTPA. The lysine dendrimer-DTPA[Gd] and avidin were conjugated in MES solution, pH 6.0, using EDC as the coupling reagent. The biotin-avidin system was used to link the polyclonal antibody and contrast agent. K562 cells were used for imaging.

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Evaluation of Antibody Immobilization Methods for Detection of Salmonella using Impedimetric Biosensor (살모넬라균 검출을 위한 임피던스 바이오센서의 항체 고정화 방법 평가)

  • Kim, Gi-Young;Moon, Ji-Hea;Om, Ae-Son;Yang, Gil-Mo;Moh, Chang-Yeon;Kang, Suk-Won;Cho, Han-Keun
    • Journal of Biosystems Engineering
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    • v.34 no.4
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    • pp.254-259
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    • 2009
  • Conventional methods for pathogen detection and identification are labor-intensive and take several days to complete. Recently developed biosensors have shown potential for the rapid detection of foodborne pathogens. In this study, an impedimetric biosensor was developed for rapid detection of Salmonella typhimurium. To develop the biosensor, an interdigitated microelectrode (IME) was fabricated by using semiconductor fabrication process. Anti-Salmonella antibodies were immobilized based on either avidin-biotin binding or self assembled monolayer (SAM) on the surface of the IME to form an active sensing layer. To evaluate effect of antibody immobilization methods on sensitivity of the sensor, detection limit of the biosensor was analyzed with Salmonella samples innoculated in phosphate buffered saline (PBS) or food extract. The impedimetric biosensor based on SAM immobilization method produced better detection limit. The biosensor could detect 107 CFU/mL of Salmonella in pork meat extract. This method may provide a simple, rapid, and sensitive method to detect foodborne pathogens.

Immunohistochemical study of the endocrine cells of the gastrointestinal mucosa of the African clawed toad, Xenopus laevis (아프리카 발톱두꺼비의 소화관내분비세포에 관한 면역조직화학적 연구)

  • Lee, Hyeung-sik;Lee, Jae-hyun
    • Korean Journal of Veterinary Research
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    • v.37 no.1
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    • pp.9-13
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    • 1997
  • The gastrointestinal endocrine cells of the African clawed toad, Xenopus laevis have been investigated immunohistochemically using the avidin-biotin method. Seven antisera were tested and three endocrine cell types immunoreacted with antisera to neurotensin, GRP and substance P. A moderate number of neurotensin-immunoreactive cells were weakly reacted in the small intestine. GRP-immunoreactive cells were mainly situated among the upper portion in the fundic glands, and the basal portion in the pyloric glands. These cells were oval and round in shape. On the other hand, in the intestine they were thin spindly cells with the epithelium. Substance P-immunoreactive cells were observed in among intestinal epithelium. However, no secretin-, motilin-, M-Enk- and PYY-immunoreactive cells were found in the GIT of the African clawed toads.

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Nutritional Requirements of Actinomyces Isolated from Rumen of Goat

  • Park, Ki Moon;Shin, Hyung Tai;Kang, Kook Hee;Lee, Jae Heung
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.1
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    • pp.61-65
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    • 2005
  • The objective of this work was to investigate the nutritional requirements for the growth of Actinomyces sp. 9RCC5 isolated from the rumen of a native goat in Korea. The growth of strain 9RCC5 on the basal medium or the medium minus certain ingredients from the basal medium demonstrated that strain 9RCC5 showed absolute requirement of vitamin B complex mixture, while hemin and volatile fatty acids (VFA) were stimulatory to growth to some extent. The 9RCC5 strain grew well with casein hydrolysate as the sole added nitrogen source. However, neither a complex of 18 amino acids nor ammonium sulfate effectively replaced casein hydrolysate. Vitamins such as riboflavin and pantothenate were essential for growth, while thiamin and biotin were stimulatory. With regard to VFA, the growth was stimulated by acetic acid but inhibited by valeric acid. Relatively large quantities of $Na^+$, $K^+$ and $Ca^{2+}$ were absolutely required for growth. Supplementation of clarified rumen fluid to the basal medium in a range of 0-10% (vol/vol) resulted in an increased rate of growth as well as an increased extent of growth.

Rapid Detection Kit for Salmonella typhimurium (살모넬라 식중독균 신속 검출용 간이 진단키트)

  • Kim, Gi-Young;Yang, Gil-Mo;Park, Saet-Byeol;Kim, Yung-Hwun;Lee, Kang-Jin;Son, Jae-Yong;Kim, Hyuck-Joo;Lee, Sae-Rom
    • Journal of Biosystems Engineering
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    • v.36 no.2
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    • pp.140-146
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    • 2011
  • This study was performed to develop a rapid test kit for pathogenic Salmonella in various samples. The rapid detection kit has been fabricated based on nitrocellulose lateral-flow strip. Colloidal gold and biotin conjugated Salmonella antibodies were used as a tag and a receptor, respectively. Manually spotted Salmonella antibody and Neutravidin on nitrocellulose membrane were used as test and control lines, respectively. Feasibility of the rapid kit to detect Salmonella typhimurium in samples were evaluated. The intensity of the color of the test line started to increase with the samples in which higher concentration of the cells were contained. The sensitivity of the sensor was $10^6$ cfu/mL Salmonella spiked in PBS. Also, the rapid test kit could detect $10^6$ cfu/mL of Salmonella in chicken meat extract.

Cerebral lesions of encephalitozoonosis in the rabbits died of rabbit hemorrhagic disease (토끼 출혈병(出血病)으로 죽은 토끼의 대뇌(大腦)에서 보인 encephalitozoonosis의 병변(病變))

  • Park, Jae-hak;Lee, Yong-soon;Itakura, C.
    • Korean Journal of Veterinary Research
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    • v.32 no.2
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    • pp.227-233
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    • 1992
  • Out of twenty rabbits which died of rabbit hemorrhagic disease spontaneously occurring in Korea, five animals had a concurrent infection with Encephalitozoon cuniculi in the cerebrum. The lesions were composed of granulomas, leptomeningitis and perivascular cuffing with mononuclear cells. The granulomas consisted of a central necrotic focus surrounded by an infiltration with plasma cells, lymphocytes and macrophages. Gliosis was associated with the granulomas. Gram-positive organisms were detected in the cerebrum from two rabbits. They were oval to rod-shaped with blunt round ends. The distribution of the pathogens was investigated by the direct avidin-biotin peroxidase complex method. They were present in pseudocysts and macrophages. Pseudocysts were found in the granulomas as well as the neuropil without cellular reactions. Some organisms were present within reticulo-endothelial cells of blood capillaries and macrophages in the subarachnoid spaces. These organisms had ultrastructural characteristics consistent with Encephalitozoon cuniculi.

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Characterization of Cell Wall Proteins from the soo1-1/ret1-1 Mutant of Saccharomyces cerevisiae

  • Lee, Dong-Won;Kim, Ki-Hyun;Chun, Se-Chul;Park, Hee-Moon
    • Journal of Microbiology
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    • v.40 no.3
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    • pp.219-223
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    • 2002
  • In order to investigate the function of Soo1p/${\alpha}$-COP during post-translational modification and intra-cellular transport of cell wall proteins in Saccharomyces cerevisiae, cell wall proteins from the soo1-1/ret1-1 mutant cells were analyzed. SDS-PAGE analysis of biotin labeled cell wall proteins suggested that the soo1-1 mutation impairs post-translational modification of cell wall proteins, such as N- and/ or Ο-glycosylation. Analysis of cell wall proteins with antibodies against ${\beta}$-1,3-glucan and ${\beta}$-1,6-glucan revealed alteration of the linkage between cell wall proteins and ${\beta}$-glucans in the soo1-1 mutant cells. Compositional sugar analysis of the cell wall proteins also suggested that the soo1-1 mutation impairs glycosylation of cell wall protein in the ER, which is crucial for the maintenance of cell wall integrity.

The Hydrolysis of Dimethyl-cis-1,3-dibenzyl-2-oxoimidazolidine-4,5-dicarboxylate by Immobilized Whole Cells of Chromobacterium chocolatum (고정화된 Chromobacterium chocolatum의 Whole Cell을 이용한 Dimethyl-cis- 1,3-dibenzyl-2-oxoimidazolidine-4,5-dicarboxylate의 가수분해)

  • Lee, Youn Jin;Shim, Sang Kyun;Ahn, Yong Hyun
    • Journal of the Korean Chemical Society
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    • v.41 no.9
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    • pp.483-487
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    • 1997
  • The whole cells of Chromobacterium chocolatum was immobilized in the matrix of polyacrylamide and then used for the hydrolysis of dimethyl-cis-1,3-dibenzyl-2-oxoimidazolidine-4,5-dicarboxylate. This hydrolysis yielded the optically active monoester ( > 96% ee) which is useful as an synthetic intermediate of (+)-biotin. We have studied the optimum condition of hydrolysis by using immobilized cells under variable concentration of substrate, reaction time and pH levels. The activity of lipase in immobilized cell was retained for longer than 4 weeks. The best conversion yield of product was obtained when 2 g of wet cell was immobilized and then reacted with 200 mg of substrate at pH 7.

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Staining patterns of squamous cell carcinomas of the larynx by monoclonal anti-cytokeratin antibodies (Monoclonal anticytokeratin antibodies 에 의한 후두편평세포암의 발현양상)

  • 도남용;전세영;이성재;최봉남
    • Proceedings of the KOR-BRONCHOESO Conference
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    • 1993.05a
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    • pp.88-88
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    • 1993
  • Immunohistochemical staining for keratin proteins may be useful as a diagnostic parameter in head and neck neoplasm. Our study evaluates the keratin antibody staining properties of normal tissues and squamous cell carcinoma of the vocal folds from surgical procedures performed on 27 cases. In normal epithelia, low molecular weight cytokeratins were strongly positive in basal layer but apparently reduced in suprabasal layers and completely negative in superficial layer. In invasive squamous cell carcinomas, low molecular weight anti-ck Ab were positive in all carcinoma cells of poorly differentiated carcinomas. On the other hand, high molecular weight anti-ck Ab were positive in almost carcinoma cells of well differentiated carcinomas.

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STABILITY OF A DISULFIDE BOND OF CHIMERIC PEPTIDE DURING IN VIVO TRANSCYTOSIS THROUGH THE BRAIN ENDOTHELIAL CELLS

  • Kang, Young-Sook;Ulrich Bickel
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1998.11a
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    • pp.150-151
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    • 1998
  • Drug delivery to the brain is facilitated by the synthesis of chimeric peptides, where in neuropharmaceuticals are linked to a vector such as an antibody to the transferrin receptor that mediates transcytosis through the blood-brain barrier (BBB). When disulfide linkers are used in the chimeric peptide, it is crucial that the S-S bridge is stable during transit and that cleavage does not occur prematurely within endothelial cells, as the peptide drug moiety would then be sequestered by the BBB instead of passing through it. The present study addressed that problem. As a model drug a metabolically stable opioid peptide, [$^3$H]DALDA (Tyr-dArg-Phe-Lys-NH$_2$), was used. It was monobiotinylated with NHS-SS-biotin to yield bio-[$^3$H]DALDA. The biotinylated peptide was bound to the vector OX26-SA which is a covalent conjugate of OX26 and streptavidin (molar ratio = 1: 1). In vitro treatment of the chimeric peptide, bio-[$^3$H]DALDA/OX26-SA, with a reducing agent, dithiothreitol, released the labeled peptide from the vector by conversion of bio-[$^3$H]DALDA to the desbiotinylated derivative, desbio-[$^3$H]DALDA.

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