• Title/Summary/Keyword: antigen analysis

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MHC Multimer: A Molecular Toolbox for Immunologists

  • Chang, Jun
    • Molecules and Cells
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    • v.44 no.5
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    • pp.328-334
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    • 2021
  • The advent of the major histocompatibility complex (MHC) multimer technology has led to a breakthrough in the quantification and analysis of antigen-specific T cells. In particular, this technology has dramatically advanced the measurement and analysis of CD8 T cells and is being applied more widely. In addition, the scope of application of MHC multimer technology is gradually expanding to other T cells such as CD4 T cells, natural killer T cells, and mucosal-associated invariant T cells. MHC multimer technology acts by complementing the T-cell receptor-MHC/peptide complex affinity, which is relatively low compared to antigen-antibody affinity, through a multivalent interaction. The application of MHC multimer technology has expanded to include various functions such as quantification and analysis of antigen-specific T cells, cell sorting, depletion, stimulation to replace antigen-presenting cells, and single-cell classification through DNA barcodes. This review aims to provide the latest knowledge of MHC multimer technology, which is constantly evolving, broaden understanding of this technology, and promote its widespread use.

Development of an Immunosensor to Detect Rat IgG Using Impedance Analyser

  • No D. H.;Kang S.;Kim G. Y.;Chung S. H.;Park Y. H.;Om A. S.;Cho S. I.
    • Agricultural and Biosystems Engineering
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    • v.5 no.1
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    • pp.21-24
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    • 2004
  • Antibody based biosensors are very selective and ultra-sensitive. Antigen-antibody reactions have been used in immunoassays. In this research, a biosensor which uses antigen-antibody reaction was developed to measure and detect rat IgG. Because the antigen-antibody reaction is a physical bounding between antigen and antibody, there are several ways to measure an antigen-antibody reaction. Among the methods, impedance analysis has short measuring time and possibilities of analyzing various properties of the reaction using frequency analysis. Rat IgG could be detected with developed biosensor and impedance analyzer. The biosensor showed good repeatability and availability of detecting concentration changes of rat IgG.

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Transient and stable expression of hepatitis B surface antigen in tomato (Lycopersicon esculentum L.)

  • Srinivas, L.;Sunil Kumar, G.B.;Ganapathi, T.R.;Revathi, C.J.;Bapat, V.A.
    • Plant Biotechnology Reports
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    • v.2 no.1
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    • pp.1-6
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    • 2008
  • Cotyledonary leaves of tomato cv. Megha were transformed with the hepatitis B virus 's' gene, which encodes surface antigen. Six plant expression cassettes (pHBS, pHER, pEFEHBS, pEFEHER, pSHER and pEFESHER) were used to assay the possible expression levels by agroinfiltration. The maximum transient expression level of 489.5 ng/g D.W. was noted in pEFEHER-infiltrated cotyledonary leaves. Transgenic tomato plants with pEFEHBS and pEFEHER expression cassettes were regenerated and characterized by molecular analysis. The expression of the antigen in the fruits was confirmed by RT-PCR and ELISA analysis. This is the first report on the expression of hepatitis B surface antigen in tomato.

Studies on antigenicity and production of monoclonal antibody for diagnosis of canine heartworm(Dirofilaria immitis) (개 심장사상충(Dirofilaria immitis) 진단을 위한 항원성 조사 및 단크론항체 생산)

  • Lee, Cheol-soon;Jee, Cha-ho
    • Korean Journal of Veterinary Research
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    • v.40 no.1
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    • pp.130-137
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    • 2000
  • In order to diagnose canine heartworm infection by antigen capture ELISA, the crude somatic(S), partial somatic(below 45kDa) and excretory/secretory(E/S) antigen of adult heartworm were identified and the antigenicity was examined by silver stain, immunoblot and ELISA. Then, production of monoclonal antibody to specific antigen carried out in this experiment. The bands to S antigen and E/S antigen were recognized between 10 and 200kDa and common bands were recognized strongly 14, 18, 28, 43kDa by silver stain. By western blot analysis, fractions to S antigen were recognized 14, 16, 18, 20, 24, 28, 32, 43, 50, 55kDa, etc. and only a 14kDa to E/S antigen in positive sera which were positive in modified Knott's test and necropsy. In ELISA, the positive sera reacted to antigens(SA, $SA_{45}$, E/S) were significantly different from negative sera by Student's t-test(p<0.05). Four hybridoma cell lines(14, 16, 17, 32kDa) than produce specific monoclonal antibodies for these antigens were obtained by immunizing BALB/c mice with a partially purified somatic antigen (below 45kDa) preparation, by fusing spleen cells with SP2/O cell myeloma cells, and by screening cell culture supernatants for antibody. In these results, it was confirmed that partial somatic antigen(below 45kDa) or E/S antigen can be used for serologic diagnosis of heartworm infection and monoclonal antibody reacting with specific antigen(14kDa) can be used for antigen capture ELISA in prepatent period of canine heartworm infection.

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Acute-onset respiratory signs in a Labrador Retriever with a positive SARS-CoV-2 rapid antigen test and infection confirmed by RT-PCR analysis: a case report

  • Mark, Gosling;Jessica, Bacon
    • Journal of Veterinary Science
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    • v.23 no.6
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    • pp.80.1-80.6
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    • 2022
  • A 10-year-old male neutered Labrador Retriever presented with a history of acute-onset tachypnoea, lethargy and anorexia. The dog was pyrexic, tachypnoeic and dyspnoeic on examination. A rapid antigen test for severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) was performed on an oropharyngeal swab and yielded a positive result. SARS-CoV-2 infection was subsequently confirmed by reverse transcription polymerase chain reaction (RT-PCR) analysis. Both of the dog's owners had positive rapid antigen test and RT-PCR analysis results for SARS-CoV-2. Additional diagnostics included computed tomography. Resolution of the dog's clinical signs was achieved with symptomatic treatment.

Immunohistochemical analysis of the effect of low power GaAlAs laser treatment on the expression of proliferating cell nuclear antigen (PCNA) in full-thickness excisional wound of rat skin (CaAlAs 저출력 레이저 자극이 흰쥐의 피부 전층결손 절제 창상의 치유시 proliferating cell nuclear antigen(PCNA)발현에 대한 면역조직화학법적 분석)

  • Kim, Soon-Ja;Koo, Hee-Seo
    • Journal of Korean Physical Therapy Science
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    • v.10 no.1
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    • pp.198-205
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    • 2003
  • We evaluated the effect of low power GaAsAl laser on re-epithelization in full-thickness excisional wound of rat skin. Two full-thickness excisions were made on the back of the experimental animals. Low power laser applications with 10mW intensity were treated experimental animals twice a day for 7 days. On the seventh postoperative day the quantitative analysis of re-epithelization was performed using immunohistochemical staining for proliferating cell nuclear antigen (PCNA). The majority of PCNA immunoreactive cells was observed at epithelial cells in the margin of full thickness excisional wound. The low power laser treatments significantly increased the number of PCNA immunoreactive cell as compared to that of non treated animal group (p<0.01). The shape of PCNA immunoreactive cell appeared as small dark, round to ovoid structures. Most PCNA immunoreactive cells exhibited a high intensity of staining that contrasted sharply with the surrounding background. In conclusion, these findings suggest that GaAlAs laser treatments effectively enhance the epithelial wound healing by the stimulating cell proliferation. Furthermore, the majority of cell proliferation occurred in the margin of full thickness excisional wound.

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Analysis of the Major Histocompatibility Complex Class I Antigen Presentation Machinery in Human Lung Cancer

  • Kim, Hyun-Pyo;Jin, Mi-Rim;Kim, Ick-Young;Ahn, Byung-Yoon;Kang, Seong-Man;Choi, Eui-Ju;Kim, Joon;Kim, Ik-Hwan;Ahn, Kwang-Seog
    • Journal of Microbiology and Biotechnology
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    • v.9 no.3
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    • pp.346-351
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    • 1999
  • Tumor cells may alter the expression of proteins involved in antigen processing and presentation, allowing them to avoid recognition and elimination by cytotoxic T cells. In order to investigate whether the major histocompatibility complex (MHC) class I-mediated antigen processing machinery is preserved in human lung cancer cell lines, we examined the expression of multiple components of the MHC class I antigen processing pathway, including transporter associated with antigen processing (TAP), $\beta_2$-microglobulin, MHC class I molecules, and chaperones which have not been previously examined in this context. Row cytometry analysis showed that the cell surface expression of MHC class I molecules was downregulated in all of the cell lines. While some cell lines showed no detectable expression of MHC class I molecules, pulse-chase experiments showed that MHC class I molecules were synthesized in the other cell lines but not transported from the endoplasmic reticulum to the cell surface. Low or nondetectable levels of TAP1 and/or TAP2 expression were demonstrated by Western blot analysis in all of the cell lines, representing a variety of lung tissue types. In some cases, this was accompanied by loss of tapasin expression. Our findings suggest that downregulation of antigen processing may be one of the strategies used by tumors to escape immune surveillance. This study provides further information for designing the potential therapeutic applications such as immunotherapy and gene therapy against cancers.

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Study on prevalence of antigens to bovine viral diarrhea virus (BVDV) of Cattle in Busan area (2013~2014) (부산지역 소 바이러스성 설사병 바이러스(bovine viral diarrhea virus; BVDV) 감염 실태 조사·연구(2013~2014))

  • Kim, Hong-Tae;Park, Min-Sik;Lee, Gi-Heun;Lee, Keun-Woo
    • Korean Journal of Veterinary Service
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    • v.38 no.1
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    • pp.43-49
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    • 2015
  • Bovine viral diarrhea virus (BVDV) is a very important viral disease virus in cattle, domestic and wild ruminants. The purpose of this study is to investigate the positive rate of bovine viral diarrhea virus antigen by ELISA from Korean native and beef cattle reared in Busan area from March in 2013 to October in 2014. A total of 1,129 bovine blood samples were collected from 140 farms, 1,111 Korean native cattle of 135 farms and 18 beef cattle of 5 farms. Test for antigen was carried out by ELISA method. In general analysis, the positive rate of bovine viral diarrhea virus antigen were 0.7% (8/1,129) cattle and 5.0% (7/140) farm. In regional analysis, the positive rate of BVDV antigen of farm in Kijang-gun, Gangseo-gu, Geumjeong-gu, Saha-gu and Dongnae-gu were 1.4% (2/94), 3.6% (5/37), 0% (0/7), 0% (0/1) and 0% (0/1), respectively, and the positive rate of BVDV antigen of cattle were 0.4% (3/770), 1.5% (5/333), 0% (0/24), 0% (0/1) and 0% (0/1), respectively. The positive rate of BVDV antigen according to sex were 0.6% (6/1,085) female cattle and 4.6% (2/44) male cattle. According to the age of cattle, the positive rate of BVDV antigen in 1 year, 2 years, 3 years and 5 years old were 1.9% (4/215), 0.4% (1/265), 0.9% (2/234) and 1.0% (1/103), respectively, but 4 years (0/198), 6 years (0/55), 7 years (0/24), 8 years (0/14), 9 years (0/10), 10 years (0/7) and 11-15 years (0/3) old were negative, respectively.

Metagonimus yokogawai: a 100-kDa Somatic Antigen Commonly Reacting with Other Trematodes

  • Han, Eun-Taek;Yang, Hyun-Jong;Park, Young-Jin;Park, Jeong-Hyun;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.52 no.2
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    • pp.201-204
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    • 2014
  • This study was undertaken to characterize the properties of a 100 kDa somatic antigen from Metagonimus yokogawai. Monoclonal antibodies (mAbs) were produced against this 100 kDa antigen, and their immunoreactivity was assessed by western blot analysis with patients' sera. The mAbs against the 100 kDa antigen commonly reacted with various kinds of trematode antigens, including intestinal (Gymnophalloides seoi), lung (Paragonimus westermani), and liver flukes (Clonorchis sinensis and Fasciola hepatica). However, this mAb showed no cross-reactions with other helminth parasites, including nematodes and cestodes. To determine the topographic distribution of the 100 kDa antigen in worm sections, indirect immunoperoxidase staining was performed. A strong positive reaction was observed in the tegumental and subtegumental layers of adult M. yokogawai and C. sinensis. The results showed that the 100 kDa somatic protein of M. yokogawai is a common antigen which recognizes a target epitope present over the tegumental layer of different trematode species.

Serological cross-reaction with Brucella abortus antigen extracted by sodium dodecyl sulfate and Yersinia enterocolitica 0:9 (SDS 처리한 브루셀라 항원과 Yersinia enterocolitica 0:9주의 혈청학적 교차반응 연구)

  • Lim, Yoon-kyu;Yang, Ki-chun;Lee, Kyung-kap;Park, Jun-hong;Lee, Du-sik;Park, Yong-ho;Kang, Seung-won;Mok, Ji-won;Lee, Yong-soon
    • Korean Journal of Veterinary Research
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    • v.35 no.1
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    • pp.143-148
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    • 1995
  • Brucella abortus cell wall antigen was extracted from Brucella abortus 1119-3 by ultrasonication and followed by sodium dodecyl sulfate(SDS) treatment. In order to confirm whether this preparation is serologically cross reactive with Yersinia enterocolitica 0:9, Western blot analysis with mouse anit-Brucella abortus1119-3 and with mouse anti-Yersinia enterocolitica 0:9 was performed. ELISA results from using those Brucella antigen and Yersinia antigen were assessed whether they had correlation. According to the results of western blot analysis and ELISA, there was no evidence of cross reactivity between the Brucella abortus 1119-3 antigen preparation and Yersinia enterocolitica 0:9. Therefore the SDS treated antigen prepared in this study could be suitably used as specific ELISA antigen without confusion in the interpretation of serological tests for brucellosis in cattle.

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