• Title/Summary/Keyword: Immunochemical Method

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Identification of Irradiated Foods by Using DNA, Immunochemical, and Biological Methods

  • Kim, Kyeung-Eun;Yang, Jae-Seung
    • Preventive Nutrition and Food Science
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    • v.4 no.4
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    • pp.276-282
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    • 1999
  • Ionizing radiation is considered to be an efficient technology to improve food safety and to extend food shelf-life in the food industry, and it has been used in food processing with a number of attributes. Food labeling should be established to enable the consumer to choose food freely, based on label information. A variety of methodologies to determine the physical, chemical, microbiological, and biological changes due to irradiation has been investigated in order to discriminate the irradiated and unirradiated food products for the consumer's free choice in food selection. However, no satisfactory method has been developed so far. In this review, various approaches based on DNA, immunochemical, and biological methods are addressed.

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Evaluation of the Spermatozoal Defect with Immunochemical Method (면역화학적 방법에 의한 정자결함 검색)

  • Kim, Se-Joong;Lee, Moo-Sang
    • Clinical and Experimental Reproductive Medicine
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    • v.18 no.1
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    • pp.101-105
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    • 1991
  • Although many therapies have been advocated in the treatment of idiopathic male infertility, the results of treatment are poor. This probably seems to be due to a lack of one or more proteins constituting the key structures of the spermatozoa. We evaluated the functional structures of the spermatozoa in 11 infertile patients whose semen showed severe oligoasthenozoospermia with immunochemical method and found a case with spermatozoa lacking acrosin. Evaluation of the spermatozoal defect with immunochemical method is desirable in patients with severe oligoasthenozoospermia, especially in cases unresponsive to medical therapy.

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Immunochemical Studies on Expression of Quinoproteins in Escherichia coli

  • Ryou, Chong-Suk;Kim, Jae-Beom;Kwon, Moo-Sik
    • Journal of Microbiology and Biotechnology
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    • v.10 no.1
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    • pp.95-98
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    • 2000
  • An immunochemical method has been develooped as the most sensitive tool for studying the expression of quinoproteins containing pyrroloquinoline qinone(PQQ) in E. coli. The PQQ was conjugated to bovine serum albumin (BSA), and the conjugant was purified by using a $KwikSep^{TM}$ dextran desalting column chromatography. The PQQ-BSA conjugant was immunized to rabbits, and the IgG fractions of the antisera were purified. The most sensitive antibody against PQQ-BSA conjugant recognized some nanogram quantity of the antigen on the blot, but had little cross reactivity with BSA. Using this batch of the antibody, all the immunochemical assays of quinoproteins in E. coli were preformed. Some six different PQQ-specfic spots were detected by Western blot analysis of the soluble proteins in E. coli were performed. Some six different PQQ-specific spots were detected by Western blot analysis of the soluble proteins in E. coli after two-dimensional gel electrophoresis. Their molecular weights on the blot were estimated to be about 100-, 90-, 72-, 58-, 52-, and 50kDa. Their pI values fell in the range from 4.8 to 5.5. These results stronly suggest that quinoproteins are present in E. coli, and that the protein moieties were covalently bound to PQQ.

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Molecular Cloning of the Gene for $\alpha$-Acylamino-$\beta$-lactam Acylhydrolase from Acetobacter turbidans by Immunochemical Detection Method (면역화학적 방법에 의한 Acetobacter turbidans의 $\alpha$-Acylamino-$\beta$-lactam Acylhydrolase의 유전자 클론화)

  • Nam, Doo-Hyun;Dewey D.Y. Ryu
    • Microbiology and Biotechnology Letters
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    • v.16 no.5
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    • pp.363-368
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    • 1988
  • Molecular cloning of gene for $\alpha$-acylamino-$\beta$-lactam acylhydrolase (ALAH) III from Acetobacter turbidans has been attempted by immunochemical detection method, in which polyclonal antibody from mouse Balb/c against this enzyme was employed as a probe. As a cloning vector, λ gtll was chosen for this purpose. Two positive clones has been selected from genomic libraries of A. turbidans, which had somewhat different binding affinities on anti-ALAH III umm and anti-$\beta$-galactosidase. By restriction analysis, both clones has been turned out to lose one of EeoRI sites. From these results, it concluded that deletion of DNA between lacZ gene and inserted DNA has occurred during replication of these clones in host cells.

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Liposome Immunoassay for Bioactive Substances (리포좀을 이용한 생리활성물질의 면역학적 분석법)

  • Kim, Chong-Kook;Park, Kyung-Mi
    • Journal of Pharmaceutical Investigation
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    • v.24 no.4
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    • pp.201-215
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    • 1994
  • Recent development in the immunochemical technique has resulted in a new ultrasensitive analytical method known as liposome immunoassay (LIA). Liposome is a key element in performing liposome immunoassays, specifically designed to participate in immune reactions. A variety of markers can be encapsulated in liposomes and used as quantitative indicators of reactions. Liposome immunoassay based on agglutination, complement-mediated Iysis, cytolysin-mediated Iysis, detergent-mediated Iysis or destabilization of the liposomal membrane have been reviewed. The quantity of markers released from liposomes should be proportional to the concentration of the analytes. Therefore, liposomal agglutination and Iysis which are essential to liposomal Iysis are critically reviewed to provide a better understanding of liposome immunoassay. Based on the literature review of recent advances in liposome immunoassay for bioactive substances, this assay method may provide a convenient, specific and highly sensitive method for detecting and measuring trace amount of clinically relevant substances in the future.

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Development of Rapid, Safe Analytical Techniques of Aflatoxins and Their Current Regulation (Aflacxin에 대한 최신 분석법과 규제동향)

  • 정덕화
    • Journal of Food Hygiene and Safety
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    • v.5 no.3
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    • pp.131-138
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    • 1990
  • Aflatoxins is a chemically diverse group of toxic secondary metabolites that are produced by fungi and often occur in agricultural commodities. Because of their wide range of toxic effects, Aflatoxins cause severe economic losses to farmers and livestock producers and pose a health to human consuming contaminated foods. Long term prospects for biotechnological control of Aflatoxins require elucidation of the specific steps and regulation of their biosynthetic pathways . Aflatoxin determinations can be approached many ways. It is essential to safely handle all experimental materials associated with aflatoxin analysis or aflatoxigenic fungi Visual screening of suspect samples, base on the presence of conidial head of the aspergillus flavus group, and screening samples for the presence of bright greenish yellow flourescence are not chemical tests and such screening techniques may allow aflactoxin contaminated lots into commerce. Microcolumn screening procedures should always be used in conjunction with a quantitative method. Several thin layer chromatography(TLC) and high performance liquid chromatography(HPLC) methods are suitable for quantitation and are in general use. Immunochemical Methods such as the ELISA or affinity column chromatography methods are being rapidly developed. The chemical and immunochemical methods can be reliable if care is taken, using suitable controls and personnel that are well trained . All analytical laboratories should stress safety and include suitable analytical validation procedure. Especially a worldwide enquiry was undertaken in recent to obtain up-to-date information about aflatoxin legislation in as many countries of the world as possible. The information concerns aflatoxin in foodstuffs. aflatoxin MI in dairy products, aflatoxins in animal feedstuffs. Limits and regulations for aflatoxin have been expended in recent with more countries having legislation on subject, more products, and more aflatoxins covered by this legislation.

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Differentitation of Yeast Species by Techniques of Electrophoresis and Immunodiffusion (단백질의 전기영동 패턴 및 항체 특성을 이용한 효모의 동정)

  • Kim, Young-Nam;Cho, Hye-Young;Kim, Joung-Han;Yoon, Suk-Kwon;Byun, Si-Myung
    • Korean Journal of Food Science and Technology
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    • v.20 no.1
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    • pp.90-94
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    • 1988
  • Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunodiffusion method were used for the species differentiation of yeasts, Saccharomyces cerevisiae, Candida utils, Candida tropicalis, and Kleuyveromyces fragilis. Comparing the electrophoretic patterns of soluble and membrane proteins, Saccharomyces cereνisiae was similar to Candida utilis but was different from Candida tropicalis and Kleuyveromyces fragilis. In immunochemical properties of soluble proteins, Saccharomyces cerevisiae was almost identical with Candido utilis. However, Saccharomyces cerevisiae or Candida utilis was quite different from Candida tropicalis and Kleuyveromyces fragilis in their immunoreactivities. In immunochemical properties of membrane proteins, almost the same results were obtained irrespective of four yeast species. By using SDS-PAGE and immunodiffusion methods, Saccharomyces cerevisiae and Candida utilis were difficult to differentiate but both species were easily differentiated from Candida tropicalis and Kleuyveromyces fragilis.

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Immunochemical Detection of Soybean Mosaic Virus Infections in the Seeds of Soybean Cultivars in Korea (면역이중확산법에 의한 콩 종자의 모자이크 바이러스(SMV) 감염상 조사)

  • La Yong-Joon;Bak Won-Chull;Oh Jeung-Haing
    • Korean journal of applied entomology
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    • v.22 no.1 s.54
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    • pp.26-30
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    • 1983
  • Soybean mosaic virus (SMV) infection in seeds wits investigated by an immunochemical method Seven soybean cultivars or lines Bughae No.1, KAS 662-7, Chungbugbaeg, Gwanggyo Clark, Bongeui, and Gangrim were tested using hypocotyls of germinated seeds and presence of SMV was detected in six soybean cultivars but Gangrim. The level of SMV infection in the assayed cultivars varied from 2.1 to $12.5\%$. It seemed that seed coat mottling had no correlation with seeds. SMV infection of the seeds since virus has not always been detected from the mot tled seeds SMV has not been detected in the seeds of variety Gwanggyo which showed necrotic symptoms.

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Noninvasive Testing for Colorectal Cancer Screening: Where Are We Now?

  • Jaeyoung Chun;Jie-Hyun Kim;Young Hoon Youn;Hyojin Park
    • Journal of Digestive Cancer Research
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    • v.11 no.2
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    • pp.85-92
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    • 2023
  • Colorectal cancer (CRC) is one of the most prevalent cancers and is the leading cause of cancer-related mortality worldwide. Based on the current screening guidelines by the American Cancer Society and Korean multi-society expert committee, CRC screening is recommended in asymptomatic adults starting at the age of 45 years. Fecal immunochemical test-based screening programs reduce the development of CRC and related mortality in the general population. However, this most popular CRC screening strategy demonstrates a crucial limitation due to modest diagnostic accuracy. Colonoscopy may be considered as an alternative primary method for CRC screening; however, its implementation can still be challenging due to concerns regarding invasiveness, low adherence, cost-effectiveness, and quality assurance. To overcome the limitations of current screening tests, innovative noninvasive tests for CRC screening have been developed with advances in molecular biology, genetics, epigenetics, and microbiomics for detecting CRC, which may enhance the approach to CRC screening and diagnosis in clinical practice in the near future. This review explores the emerging screening methods and discusses their potential for integration into current practice.

Immunochemical Studies of Starfish Gangliosides: Production of Monoclonal Antibody against AG-2, the Major Ganglioside of Starfish Acanthaster planci, and Detecting Its Distribution in Tissues by TLC Immunostaining

  • Miyamoto, Tomofumi;Yamamoto, Atsushi;Sakai, Maki;Tanaka, Hiroyuki;Shoyama, Yukihiro;Higuchi, Ryuichi
    • Journal of Marine Bioscience and Biotechnology
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    • v.1 no.4
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    • pp.298-304
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    • 2006
  • In this study, we establish a thin-layer chromatography (TLC) immunostaining method for detecting starfish gangliosides. A new monoclonal antibody (MAb) against AG-2, the major gangliosides molecular species of Acanthaster planci, was produced by fusing hybridoma with splenocytes immunized to liposomal AG-2. BALB/c male mice were injected with liposomal AG-2 antigen, and immunized. Their splenocytos were isolated and fused with hypoxanthine-aminopterine-thimidine (HAT)-sensitive mouse myeloma cells. Hybridomas producing MAb reactive to AG-2 were cloned using the limited dilution method. Established hybridomas were cultured in eRDF medium. Crude MAb produced from clone 8D4 was purified with a magnesium pyrophosphate column. Enzyme immunoassay and TLC immunostaining of AG-2 were performed using the purified MAb. Structurally related gangliosides did not cross-react with anti-AG-2 antibodies. The detection limit of TLC immunostaining was 50 ng of AG-2. The newly established immunostaining method was further developed for detecting AG-2 distribution and qualitative analysis in tissues and/or organs. Our results show that the majority of AG-2 is present in the stomach of male A. planci, while AG-2 is distributed not only in the stomach but also in the the pyloric caeca of female A. planci.

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