• Title/Summary/Keyword: specific labeling

Search Result 208, Processing Time 0.029 seconds

A study on the body fluid antigen of Clonorchis sinensis using immunogold labeling method (면역황금 표기법을 이용한 간흡충의 체액 항원에 관한 연구)

  • Ju, Bong-Deok;Im, Han-Jong;Kim, Su-Jin
    • Parasites, Hosts and Diseases
    • /
    • v.28 no.1
    • /
    • pp.11-24
    • /
    • 1990
  • In order to observe the antigenic localization in the tissues of the adult Clonorchis sinensis, immunogold labeling method was applied using serum immunoglobulins (IgG) of either worm·infected rabbits (group I) or antigen-immunized rabbits (group II) (by the body quid obtained from the adult worms). The electron micrographs of the sectioned worm tissue antigens, embedded in Lowicryl HM 20 medium and stained with protein A-gold complex (particle sixte: 12 nm), were compared between the group I and group II. The gold particles were observed in the interstitial matrix of the worm parenchyma, the epithelial lamellae of the cecum, and the cecal lumen both in group I and II. But the particles were in general more concentrated in group II. The gold particles were not observed on the basal lamina of the tegument or on vitelline glands in group I, while they were highly concentrated on those areas in group II. There were also differences in the antigenicity of interstitial matrix(reacted with group I IgG) and head part(reacted with group II IgG) of the sperm cells in the seminal receptacle. Conclusively, it is suggested that the substances comprising the basal lamina of the tegument or vitelline glands act as specific antigens reacting with antigen(body quid) immunized rabbit IgG. On the other hand, the substances in the cecal lumen and cecal epithelial lamellae are thought to be the specific antigen that react with the worm-infected rabbit IgG.

  • PDF

Development of Species-Specific PCR Primers for the Rapid and Simultaneous Identification of the Six Species of Genus Takifugu

  • Dong, Chun Mae;Park, Yeon Jung;Noh, Jae Koo;Noh, Eun Soo;An, Cheul Min;Kang, Jung-Ha;Park, Jung Youn;Kim, Eun-Mi
    • Development and Reproduction
    • /
    • v.23 no.4
    • /
    • pp.367-375
    • /
    • 2019
  • Pufferfish (Takifugu spp.) are economically important edible marine fish. Mistakes in pufferfish classification can lead to poisoning; therefore, accurate species identification is critical. In this study, we used the mtDNA cytochrome c oxidase subunit I gene (COI) to design specific primers for six Takifugu species among the 21 domestic or imported pufferfish species legally sold for consumption in Korea. We rapidly and simultaneously identified these pufferfish species using a highly efficient, multiplex polymerase chain reaction (PCR) system with the six species-specific primers. The results showed that species-specific multiplex PCR (multiplex species-specific polymerase chain reaction; MSS-PCR) either specifically amplified PCR products of a unique size or failed. MSS-PCR yielded amplification fragment lengths of 897 bp for Takifugu pardalis, 822 bp for T. porphyreus, 667 bp for T. niphobles, 454 bp for T. poecilonotus, 366 bp for T. rubripes, and 230 bp for T. xanthpterus using the species-specific primers and a control primer (ca. 1,200 bp). We visualized the results using agarose gel electrophoresis to obtain accurate contrasts of the six Takifugu species. MSS-PCR analysis is easily performed and provides identification results within 6 h. This technique is a powerful tool for the discrimination of Takifugu species and will help prevent falsified labeling, protect consumer rights, and reduce the risk of pufferfish poisoning..

A Probing of Inhibition Effect on Specific Interaction Between Glucose Ligand Carrying Polymer and HepG2 Cells

  • Park, Keun-Hong;Park, Sang-Hyug;Lee, Hyun-Jung;Min, Byoung-Hyun
    • Journal of Microbiology and Biotechnology
    • /
    • v.14 no.3
    • /
    • pp.450-455
    • /
    • 2004
  • A reducing glucose-carrying polymer, called poly [3-O-(4'-vinylbenzyl)-D-glucose](PVG), was interacted with HepG2 cells including a type-l glucose transporter (GLUT-1) on the cell membrane. The cooperative interaction between a number of GLUT-1s and a number of reducing 3-O-methyl-D-glucose moieties on the PVG polymer chain was found to be responsible for the increase in the interaction with HepG2 cells. The affinity between the cells and the PVG was studied using RITC-labeled glycopolymers. The specific interaction between the GLUT-1 on HepG2 cells and the PVG polymer carrying reducing glucose moieties was suppressed by the inhibitors, phloretin, phloridzin, and cytochalasin B. Direct observation by confocal laser microscopy with the use of RITC-labeled PVG and pretreatment of HepG2 cells with the inhibitors demonstrated that the cells interacted with the soluble form of the PVG polymer via GLUT-1, while fluorescence labeling of the cell surface was prevented after pretreatment with the inhibitors of GLUT-1.

The Importance of FACS Analysis in the Development of Aptamers Specific to Pathogens

  • Moon, Ji-Hea;Kim, Giyoung;Park, Saet Byeol;Lim, Jongguk;Mo, Changyeun
    • Journal of Biosystems Engineering
    • /
    • v.39 no.2
    • /
    • pp.111-114
    • /
    • 2014
  • Purpose: This review aims to introduce aptamers and the methods of its development to improve the sensitivity and selectivity to target bacteria. In this review, we have highlighted current developments and directions in the pathogen detection based on aptamers. Background: Aptamers, the specific nucleic acid sequences, can bind to targets with high affinity and specificity. Some of researches on the use of aptamers for the detection of pathogen have been reported in recent years. Aptamers have more applicability than antibodies for the development of pathogen detection using biosensor; such as easy to synthesis and labeling, lack of immunogenicity, and a low cost of production. However, only few reports on the development and use of aptamers for the detection of pathogen have been published. Review: Aptamers specific to pathogen are obtained by whole-cell systematic evolution of ligands by exponential enrichment (SELEX) process. SELEX process is composed of screening random oligonucleotide bound with target cells, multiple separation and amplification of nucleic acids, final identification of the best sequences. For improving those affinity and selectivity to target bacteria, optimization of multiple separating process to remove unbounded oligonucleotides from aptamer candidates and sorting process by flow cytometry are required.

NMR study of the interaction of T4 Endonuclease V with DNA

  • Lee, Bong-Jin;Im, Hoo-Kang;Hyungmi Lihm;Yu, Jun-Suk
    • Proceedings of the Korean Society of Applied Pharmacology
    • /
    • 1995.04a
    • /
    • pp.80-80
    • /
    • 1995
  • T4 Endonuclease V (Mw 16,000) acts as a repair enzyme for UV induced pyrimidine dimers in DNA. Many researchers have studied the biochemical characteristics of the enzyme. However the precise action mechanism of T4 endo V has not fully elucidated yet. In our laboratory NMR spectroscopy technique is being used for the structural study of T4 endo V. Because of its low temperature stability and high content of ${\alpha}$-helix, the conventional $^1$H NMR technique was inapplicable. Therefore we utilized stable isotope labeling technique and so far prepared about 10 amino acid specific labeled proteins. The HSQC spectra of amino acid specific labeled proteins will help us to interpret the triple resonance 3D, 4D data which are under processing, We also studied the behaviors of specific amino acid residues whose roles might be critical. When the enzyme labeled by $\^$15/N-Thr was mixed with the substrate oligonucleotide (semispecific -TT- sequence), one crosspeak in its HSQC spectrum was completely desappeared, which means that one of seven Thr residues is in the binding site of the enzyme with DNA, This result is well consistent with previous report that implicated the Thr 2 residue in the activity of the enzyme. Similar studies were carried on the behaviors of Arg and Tyr residues.

  • PDF

Radioimmunotherapy (II): Clinical Application (방사면역치료(II): 임상적 이용)

  • Cheon, Gi-Jeong;Kang, Hye-Jin;Lim, Sang-Moo
    • Nuclear Medicine and Molecular Imaging
    • /
    • v.40 no.2
    • /
    • pp.74-81
    • /
    • 2006
  • Molecular targeting may be defined as the specific concentration of a diagnostic or therapeutic tracer by its Interaction with a molecular species that is distinctly present or absent in a disease state. Monoclonal antibody (mAb) is one of the successful agents for targeted therapy in cancer. To enhance the therapeutic effect, the concept of targeting radionuclides to tumors using radiolabeled mAbs against tumor-associated antigens, radioimmunotherapy, was proposed. The efficacy of radioimmunotherapy, however, has to be further optimized. Several strategies to improve targeting of tumors with radiolabeled mAbs have been developed, such as the use of mAb fragments, the use of high-affinity mAbs, the use of labeling techniques that are stable in vivo, active removal of the radiolabeled mAb from the circulation, and pretargeting strategies. Until now, however, there are many kinds of obstacles to be solved in the use of mAb for the targeted therapy. Major technical challenges to molecular targeting are related to the rapid and specific delivery of tracers to the target, the elimination of unwanted background activity, and the development of more specific targets to create a cytocidal effect. further development of this field will be determined by success in solving these challenges.

$^{99m}Tc-Labeling$ of Monoclonal Antibody to Carcinoembryonic Antigen and Biodistribution (항 암태아성항원에 대한 단세포군항체의 $^{99m}Tc$ 표지법개발 및 생체분포)

  • Moon, Dae-Hyuk;Chung, June-Key;Lee, Myung-Chul;Koh, Chang-Soon;Chung, Hong-Keun;Park, Jae-Gahb
    • The Korean Journal of Nuclear Medicine
    • /
    • v.26 no.2
    • /
    • pp.380-391
    • /
    • 1992
  • This study was designed to evaluate a direct method of $^{99m}Tc$ labeling using $\beta-mercaptoethanol$ as a reducing agent, and to investigate whether $^{99m}Tc$ labeled specific monoclonal antibody against carcinoembryonic antigen (CEA-92) can be used for the scintigraphic localization of human colon cancer xenograft. Purified CEA-92 IgG was fragmented into F $(ab')_2$ and then labeled with $^{99m}Tc$ by transchelation method using glucarate as a chelator. Labeling efficiency, immunological reactivity and in vitro stability of $^{99m}Tc$ CEA-92 F $(ab')_2$ were measured and then injected intravenously into nude mice bearing human colon cancer (SNU-C4). Scintigrams were obtained at 24 hour after injection. Then nude mice were sacrificed and the radioactivity was measured Labeling efficiency of injected $^{99m}Tc$ CEA-92 F $(ab')_2$, immunoreative fraction and in vitro stability at 24 hour of injected $^{99m}Tc$ CEA-92 F $(ab')_2$ was 45.2%, 32.8% and 57.4%, respectively. At 24 hour after injection, % ID/g in kidney (46.77) showed high uptake, but %ID/g in tumor (1.65) was significantly higher than spleen (0.69), muscle (0.16), intestine (0.45), stomach (0.75), heart (0.48) and blood (0.45). There was no significant difference between tumor and liver (1.81). Tumor contrast as quantitated by tumor to blood ratio of $^{99m}Tc$ CEA-92 F $(ab')_2$ was increased significantly (p<0.005) until 24 hours (3.70), and there was no statistical differece from tumor to blood ratio of I-131 CEA-92 F $(ab')_2$. The scintigram demonstrated localization of radioactivity over transplanted tumor, but significant background radioactivity was also noted over kidney and abdomen. It is concluded that CEA-92 F $(ab')_2$ can be labeled with $^{99m}Tc$ by a direct transchelation method using $\beta-mercaptoethanol$ as a reducing agent and $^{99m}Tc$ labeled CEA-92 F $(ab')_2$ can be used for the scintigraphic localization of human colon cancer xenograft in nude mice model.

  • PDF

A Survey on the Actual Condition of Products not Labeled with Allergens (알레르기 유발물질 미표시 제품 실태 조사)

  • Kim, Kyung-Seon;Song, Sung-Min;Kwon, Sung-Hee;Jang, Seung-Eun;Lee, Bo-Min;Kim, Meyong-Hee;Han, Young-Sun;Hur, Myung-Je;Kwon, Mun-Ju
    • Journal of Food Hygiene and Safety
    • /
    • v.36 no.3
    • /
    • pp.257-263
    • /
    • 2021
  • For this survey, PCR (polymerase chain reaction) testing was conducted using 14 species-specific primers to monitor the labeling of allergy-causing substances in various foods. Sixty samples from stationary stores near elementary schools and imported confectionery shops were tested, including snacks, candies, and chocolate. Allergens of milk, wheat, eggs, tomatoes, almonds and peanuts were detected in 30 cases (50.0%). In addition, many products were detected as either containing unlabeled substances or not showing allergen-related information and labeling in Korean. In order to ensure that consumers are able to purchase products safely and securely, a system for thorough guidance and monitoring of allergen-related labeling by domestic manufacturing and processing companies and import-related companies is required.

Identification of Korean Native Pork Using Breed-Specific DNA Marker of KIT Gene

  • Chung, Eui-Ryong;Chung, Ku-Young
    • Food Science of Animal Resources
    • /
    • v.30 no.3
    • /
    • pp.403-409
    • /
    • 2010
  • Accurate methods for the identification of closely related species or breeds in raw and processed meats must be developed in order to protect both consumers and producers from mislabeling and fraud. This paper describes the development of DNA markers for the discrimination and improvement of Korean native pig (KNP) meat. The KIT gene is related to pig coat color and is often used as a candidate marker. A 538 bp fragment comprising intron 19 of the pig KIT gene was amplified by PCR using specific primers, after which the PCR amplicons of a number of meat samples from KNP and three major improved breeds (Landrace, Duroc and Yorkshire) were sequenced in order to find a nucleotide region suitable for PCR-RFLP analysis. Sequence data showed the presence of two nucleotide substitutions, g.276G>A and g.295A>C, between KNP and the improved pig breeds. Digestion of KIT amplicons with AccII enzyme generated characteristic PCR-RFLP profiles that allowed discrimination between meats from KNP and improved pig. KNP showed three visible DNA bands of 264/249, 199, and 75 bp, whereas DNA bands of 249, 199, and 90 bp were detected in the three improved pig breeds. Therefore, the 75 bp DNA fragment was specific only to KNP, whereas the 90 bp DNA fragment was specific to the improved breeds. The breed-specific DNA markers reported here that target the KIT gene could be useful for the identification of KNP meat from improved pig meats, thus contributing to the prevention of falsified breed labeling.

Rapid Origin Determination of the Northern Mauxia Shrimp (Acetes chinensis) Based on Allele Specific Polymerase Chain Reaction of Partial Mitochondrial 16S rRNA Gene

  • Kang, Jung-Ha;Noh, Eun-Soo;Park, Jung-Youn;An, Chel-Min;Choi, Jung-Hwa;Kim, Jin-Koo
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.28 no.4
    • /
    • pp.568-572
    • /
    • 2015
  • Acetes chinensis is an economically important shrimp that belongs to the Sergestidae family; following fermentation, A. chinensis' economic value, however, is low in China, and much of the catch in China is exported to Korea at a low price, thus leading to potential false labeling. For this reason, we developed a simple method to identify A. chinensis' origin using allele-specific polymerase chain reaction (PCR). Ten single nucleotide polymorphisms (SNPs) were identified from partial (i.e., 570 bp) DNA sequence analysis of the mitochondrial 16s rRNA gene in 96 Korean and 96 Chinese individual shrimp. Among 10 SNP sites, four sites were observed in populations from both countries, and two sites located in the middle with SNP sites at their 3'-ends were used to design allele-specific primers. Among the eight internal primers, the C220F primer specific to the Chinese A. chinensis population amplified a DNA fragment of 364 bp only from that population. We were able to identify the A. chinensis population origin with 100% accuracy using multiplex PCR performed with two external primers and C220F primers. These results show that the 16S rRNA gene that is generally used for the identification of species can be used for the identification of the origin within species of A. chinensis, which is an important finding for the fair trade of the species between Korea and China.