• Title/Summary/Keyword: ELISA antibody

Search Result 749, Processing Time 0.032 seconds

Changes in the Titer of Tooth Root Antibodies Accompanying Root Resorption Associated with Orthodontic Tooth Movement (치아이동시 치근 흡수에 따른 치근항체의 역가 변화)

  • Park, Soo-Byung;Son, Woo-Sung
    • The korean journal of orthodontics
    • /
    • v.24 no.2
    • /
    • pp.303-317
    • /
    • 1994
  • This study was designed to measure the changes in the titer of tooth root antibodies accompanying root resorption associated with orthodontic tooth movement in dogs to explore a role of the specific immune response in root resorption during orthodontic tooth movement. Five adult mongrel dogs, 2 years of age, were used in the study. Six lower incisors were extracted as sources of homologous antigen in the dogs. Tooth root antigen preparations were made from a 6M Guanidine-HCl-10% EDTA(pH5.0) extract of these root dentins. Root resorption was elicited by intrusion of six maxillary incisors with 200-250gm intrusive force. In 9th week, resorbing six maxillary anterior teeth were extracted. Serum samples were taken from each dog prior to intrusion and weekly for 11 consecutive weeks. Serum autoantibody titers were determined with an enzyme-linked immunosorbent assay. As controls for antibody specificity, sera which were previously incubated with tooth root antigen as well as sera to an unrelated bacterial antigen (Porphyromonas gingivalis 33277) for 3 hours at 25 were measured in all runs. Root resorption was monitored monthly using occlusal radiographs. And then root resorption patterns were observed with a zoom stereo microscope (Model SZH-121, Olympus optical Co. Ltd.). Incisors did not show clear radiographic evidence of significant and progressive root resorption, but periodontal ligament space had widened. But root resorption was observed on the apical regions of the maxillary incisors with a zoom stereo microscope. Teeth showed the shallow depression generally accompanying deep resorption. These demonstrate a slight tendency for an immediate decrease followed by rebound to levels above the pre-treatment baseline. A peak titer of autoantibody to dentin antigen occurred on day 28, then steadily decreased during the 9th week period as the roots resorbed and then rapidly spiked in animals when the resorbing teeth were extracted. When sera is incubated with tooth root antigen, serum activity in the ELISA was almost absent. This is because serum activity in the ELISA could be removed by absorption of the serum with dog dentin antigen. Serum ELISA activity to the unrelated bacterial antigen remained essentially unchanged in all animals throughout the experimental period. When the time course of changes in autoantibody to homologous tooth root antigen prepatration and unrelated bacterial antigen was compared, no significant differences were found(${\alpha}=0.05$). In general, the overall pattern of changes in autoantibody was similar to the two antigens. These findings suggest the possibility that these immunologic changes precede a significant development of root resorption lesions rather than merely reflecting their presence. Therefore, this suggests that the changes of antibody levels may have some predictive value for root resorption.

  • PDF

Characterization of a Monoclonal Antibody Specific to Human Siah-1 Interacting Protein (인체 SIP 단백질에 특이적인 단일클론 항체의 특성)

  • Yoon, Sun Young;Joo, Jong Hyuck;Kim, Joo Heon;Kang, Ho Bum;Kim, Jin Sook;Lee, Younghee;Kwon, Do Hwan;Kim, Chang Nam;Choe, In Seong;Kim, Jae Wha
    • IMMUNE NETWORK
    • /
    • v.4 no.1
    • /
    • pp.23-30
    • /
    • 2004
  • Background: A human orthologue of mouse S100A6-binding protein (CacyBP), Siah-1-interacting protein (SIP) had been shown to be a component of novel ubiquitinylation pathway regulating $\beta$-catenin degradation. The role of the protein seems to be important in cell proliferation and cancer evolution but the expression pattern of SIP in actively dividing cancer tissues has not been known. For the elucidation of the role of SIP protein in carcinogenesis, it is essential to produce monoclonal antibodies specific to the protein. Methods: cDNA sequence coding for ORF region of human SIP gene was amplified and cloned into an expression vector to produce His-tag fusion protein. Recombinant SIP protein and monoclonal antibody to the protein were produced. The N-terminal specificity of anti-SIP monoclonal antibody was conformed by immunoblot analysis and enzyme linked immunosorbent assay (ELISA). To study the relation between SIP and colon carcinogenesis, the presence of SIP protein in colon carcinoma tissues was visualized by immunostaining using the monoclonal antibody produced in this study. Results: His-tag-SIP (NSIP) recombinant protein was produced and purified. A monoclonal antibody (Korea patent pending; #2003-45296) to the protein was produced and employed to analyze the expression pattern of SIP in colon carcinoma tissues. Conclusion: The data suggested that anti-SIP monoclonal antibody produced here was valuable for the diagnosis of colon carcinoma and elucidation of the mechanism of colon carcinogenesis.

The Association of HLA-DRB1 and DQB1 Alleles and a Study of Anticardiolipin Antibody and Anti-β2 Glycoprotein I Antibody in Korean SLE Patients (한국인 전신성홍반성루푸스 환자에서 HLA-DRB1, DQB1 대립유전자의 연관성 및 항인지질 항체와 항β2 Glycoprotein I 항체에 관한 연구)

  • Lee, Sang Gon;Cha, Hoon Suk;Yang, Yoon Sun
    • IMMUNE NETWORK
    • /
    • v.2 no.4
    • /
    • pp.227-232
    • /
    • 2002
  • Background: Systemic lupus erythematosus (SLE) is a complex autoimmune disease characterized by diverse clinical manifestations and autoantibody production, which is known to be strongly influenced by genetic factors. Previous studies have revealed the associations of SLE with HLA class II alleles and antiphospholipid antibody system (anticardiolipin antibody (aCL) and anti-${\beta}_2$ glycoprotein I antibody (anti-${\beta}_2$ GPI)). Therefore, we studied the associations of HLA class II alleles with SLE and antiphospholipid antibody system. Methods: The genotyping for HLA-DRB1 and DQB1 alleles were performed in 61 SLE patients and 100 controls by the polymerase chain reaction (PCR)-sequence specific oligonucleotide probe method. ELISA tests for aCL and anti-${\beta}_2$ GPI were performed in 39 of the 61 SLE patients. The results were evaluated statistically by Chi-square test. Results: The frequencies of the HLA-$DRB1^*15$ and $DQB1^*06$ in SLE patients were significantly higher than those in controls. HLA-$DRB1^*12$ was significantly lower in SLE patients than controls. Nine of 39 patients were positive for aCL (IgG) and three were positive for aCL (IgM). One of 39 patients were positive for anti-${\beta}_2$ GPI (IgG) and none of them positive for anti-${\beta}_2$ GPI (IgM). Association of aCL with HLA class II alleles was not observed in our study. Conclusion: According to our results, it was found that HLA-$DRB1^*15$ and $DQB1^*06$ were associated with genetic susceptiblility and $DRB1^*12$ was associated with resistance to SLE in Korean population. No Association of aCL with HLA class II alleles was observed and the positive rate for anti-${\beta}_2$ GPI was very low.

Quantitative Analysis of Microcystins, Cyanobacterial Toxins in Soyang Lake (소양호에서 남조류 독소, 마이크로시틴의 정량 분석)

  • Lee, Jeong Ae;Lee, So Yeong;Pyo, Dong Jin
    • Journal of the Korean Chemical Society
    • /
    • v.46 no.6
    • /
    • pp.535-540
    • /
    • 2002
  • It is very difficult to analyze the microcystins, cyanobacterial toxins quantitatively since it exists in a trace level in lakes. In this paper, two different analytical methods were tried to analyze the microcystins, cyanobacterial toxins quantitatively in water samples collected in Soyang lake. The first method was solid phase extraction method fol-lowed by High Performance Liquid Chromatography(HPLC), and the second method was Enzyme-Linked Immu-nosorbent Assay(ELISA) using the monoclonal antibody of microcystin.

Dietary Exposure of Aflatoxin$B_1$ and Cancer Risk Assessment (아플라톡식 $B_1$ 노출에 의한 발암 위해성 평가)

  • 이병무;최문정;변수현;김형식
    • Journal of Food Hygiene and Safety
    • /
    • v.10 no.2
    • /
    • pp.81-87
    • /
    • 1995
  • Daily exposure of aflatoxin B1 (AFB1) was estimated in foods (rice, barley, soybean, peanut, soysauce, soybean paste) by ELISA (enzyme linked immunosorbent assay) using polyclonal antibody R101. Before ELISA, a simple extraction method was applied for the quantitation of AFB1 in foods using chloroform which showed high recovery (70$\pm$12%). AFB1 levels in foods were 0.32 ng/ml (rice), 0.24ng/ml (barley), 0.22 ng/ml (peanut), 0.30~0.78 ng/ml (soysauce), and 0.2 ng/ml (soybean paste). Based on food consumption, we estimated that Koreans were exposed to AFB1 at the level of 1.86$\pm$0.46 ng/kg/day and liver cancer incidence attributed to AFB1 exposure (assuming that AFB1 as a single hepatocarcinogenic agent) might be calculated to be 13.1 per 100, 000 population. Our data demonstrate that AFB1 levels in foods were below the regulation of 10 ppb in foods and might not be the major risk factor for the high incidence of lover cancer in Korea.

  • PDF

Development of Recombinant Coat Protein Antibody Based IC-RT-PCR and Comparison of its Sensitivity with Other Immunoassays for the Detection of Papaya Ringspot Virus Isolates from India

  • Sreenivasulu, M.;Gopal, D.V.R. Sai
    • The Plant Pathology Journal
    • /
    • v.26 no.1
    • /
    • pp.25-31
    • /
    • 2010
  • Papaya ringspot virus (PRSV) causes the most widespread and devastating disease in papaya. Isolates of PRSV originating from different geographical regions in south India were collected and maintained on natural host papaya. The entire coat protein (CP) gene of Papaya ringspot virus-P biotype (PRSV-P) was amplified by RTPCR. The amplicon was inserted into pGEM-T vector, sequenced and sub cloned into a bacterial expression vector pRSET-A using a directional cloning strategy. The PRSV coat protein was over-expressed as a fusion protein in Escherichia coli. SDS-PAGE gel revealed that CP expressed as a ~40 kDa protein. The recombinant coat protein (rCP) fused with 6x His-tag was purified from E.coli using Ni-NTA resin. The antigenicity of the fusion protein was determined by western blot analysis using antibodies raised against purified PRSV. The purified rCP was used as an antigen to produce high titer PRSV specific polyclonal antiserum. The resulting antiserum was used to develop an immunocapture reverse transcription-polymerase chain reaction (IC-RT-PCR) assay and compared its sensitivity levels with ELISA based assays for detection of PRSV isolates. IC-RT-PCR was shown to be the most sensitive test followed by dot-blot immunobinding assay (DBIA) and plate trapped ELISA.

Enzymatic N-glycan analysis of 31 kDa molecule in plerocercoid of Spirometra mansoni (sparganum) and its antigenicity after chemical oxidation

  • Chung, Young-Bae;Kong, Yoon;Yang, Hyun-Jong
    • Parasites, Hosts and Diseases
    • /
    • v.42 no.2
    • /
    • pp.57-60
    • /
    • 2004
  • A highly specific antigenic protein of 31 kDa from plerocercoid of Spirometra mansoni (sparganum) was obtained by gelatin affinity and Mono Q anion-exchange column chromatography. The purified 31 kDa protein was subjected to N-glycan enzymatic digestion for structural analysis. The relative electrophoretic mobility was analyzed by SDS-PAGE, before and after digestion. On SDS-PAGE after enzymatic digestion, the 31 kDa protein showed a molecular shift of approximately 2 kDa, which indicated the possession of complex N-linked oligosaccharides (N-glycosidase F sensitive) but not of high-mannose oligosaccharides (endo-beta-N-acetylglucosaminidase H, non-sensitive). Chemically periodated 31 kDa protein showed statistically non-significant changes with human sparganosis sera by enzyme linked immunosorbent assay (ELISA). Therefore, the dominant epitopes of the 31 kDa molecule in human sparganosis were found to be mainly polypeptide, while N-glycans of the antigenic molecule in sparganum was minimal in anti-carbohydrate antibody production.

Examination of seroprevalence and detection of Avian pneumovirus from layer hens in Gyeongbuk province (경북지역 산란계에서 Avian pneumovirus에 대한 항체가 및 바이러스 검출 조사)

  • Kim, Jung-Eun;Hwang, Ji-Young;Bae, Dong-Rok;Sung, Myoung-Suk;Kim, Soon-Tae;Kim, Sang-Yun
    • Korean Journal of Veterinary Service
    • /
    • v.30 no.1
    • /
    • pp.43-49
    • /
    • 2007
  • Avian pneumovirus(APV), also known as avian rhinotracheitis(ARTV), affects both turkeys and chickens and is known to be the primary causative agent of turkey rhinotracheitis(TRT). The aim of this study was to establish the presence or absence of antibodies to APV by an enzyme-linked immunosorbent assay(ELISA) and confirm APV by a reverse transcriptase-polymerase chain reaction(RT-PCR). The tested serum and feces were collected from laying hens in Gyeongbuk province. The positive farms with antibody against APV by ELISA were 90(96.7%) of 93 and positive serum samples were 433(93.1%) of 465 different sera. By regional group, sera from Uiseong, Cheongsong and Bonghwa were noted as 100% positive and positive rates of samples from Yeongju, Andong and Yeongyang were 93.3%, 85.7% and 50%, respectively. However, APV was not detected in feces samples by RT-PCR.

Production of Monoclonal Antibodies Against Nervous Necrosis Virus (NNV, RGNNV genotype) (신경괴사증바이러스(nervous necrosis virus, RGNNV genotype)에 대한 단클론 항체 생산)

  • Kim, Wi-Sik;Kim, Si-Woo;Oh, Myung-Joo
    • Korean Journal of Fisheries and Aquatic Sciences
    • /
    • v.51 no.3
    • /
    • pp.328-331
    • /
    • 2018
  • We developed and subsequently characterized mouse monoclonal antibodies (MAbs) against nervous necrosis virus (NNV, RGNNV genotype). We established six hybridoma clones secreting MAbs against NNV antigen: 2B1, 2B11, 2C12, 13C1-1, 13C1-2 and 14D11. All six MAbs belonged to the IgG2a isotype with a kappa light chain and their reactivity recognized against the 41 kDa coat protein of NNV by Western blot analysis. The affinity constants of the six MAbs were measured by enzyme-linked immunosorbent assay (ELISA). All six MAbs reacted with two NNV isolates (SgNag05 and Gemunodo06), while no reactivity was observed with five know fish viruses, namely marine birnavirus, infectious pancreatic necrosis virus, viral hemorrhagic septicemia virus, hirame rhabdovirus, and infectious hematopoietic necrosis virus. Moreover, high ELISA optical density (OD) values (0.87-1.42) were observed in the brain tissues of NNV-infected sevenband grouper, while low OD values (less than 0.12) were recorded in the brain tissues of uninfected fish. These results suggest that these six MAbs are highly competent and useful for the detection of NNV with the RGNNV genotype.

Serotaxonomical Analyses of Some Streptomyces Strains Using Antibodies against Cell Envelope (균사 외피 항체를 이용한 Streptomyces 속 균주들의 혈청학적 유사성 분석)

  • Jo, Sung-Kee;Kim, Jae-Heon
    • Korean Journal of Microbiology
    • /
    • v.43 no.2
    • /
    • pp.137-141
    • /
    • 2007
  • The similarities among five strains of Streptomyces were measured by the serological methods. Antigens were prepared by dissolving cell envelope fractions in Tween 20 buffer and antisera were produced from the immunized rabbits. Immunodiffusion studies and ELISA results showed that the degree of antigen-antibody reaction was not exactly matched to the taxonomic distance; i. e. the strains of the cluster group A exhibited low level of cross-reactions each other, while relatively strong cross-reactions were observed between Streptomyces lavendulae of cluster group F and Streptomyces viridochromogenes of cluster group A.