• Title/Summary/Keyword: Antibody screening

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Meaning of Anti Hepatitis C Virus Antibody Detection in Patients Treated with Intravenous Immunoglobulin (인민역 글로불린 제재를 투여받은 환아에서 C형 간염바이러스 항체(anti-HCV) 검출의 의의)

  • Lee, Myong-Gyun;Lee, Young-Hwan;Shin, Son-Moon;Hah, Jeong-Ok
    • Journal of Yeungnam Medical Science
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    • v.10 no.2
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    • pp.306-312
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    • 1993
  • To evaluate the meaning of anti-HCV detection in patients treated with IVIG, serum levels of aspartate aminotranstferase(AST), alanine aminotransterase(ALT), HCV Ab titer were measured after treatment with IVIG in 36 patients diagnised of Kawasaki disease or neonatal sepsis. Also polymerase chain reaction (PCR) for the detection of HCV was done in 8 patients with persistent HCV Ab positivity at 3 months after IVIG treatment. The results were as follows 1) HCV Ab was positive in all 36 patients at 1 week after IVIG treatment, but in only 8 cases it was positive at 3 months after IVIG treatment. 2) AST, ALT were elevated in 9 cases at 1 week after IVIG treatment, but they were normalized in all cases at 3 months after IVIG treatment. 3) PCR for the detection of HCV was done in 8 patients with persistent HCV Ab positivity at 3 months after IVIG treatment, but HCV was not isolated in any cases. These results suggested that detection of anti-HCV was merely transitory phenominon of HCV Ab transmission, did not show any evidence of HCV infection due to HCV transmission.

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Studies on the Immunoblot Characterization of Clonorchis sinensis Worm Antigens at Carly Development Stages (Immunoblot 법을 이용한 간흡충항원(肝吸蟲抗原)의 발육단계별(發育段階別) 항원성분석(抗原性分析)에 관한 연구(硏究))

  • Lee, Seon-Kyung;Joo, Kyoung-Hwan;Chung, Myung-Sook;Rim, Han-Jong
    • Journal of agricultural medicine and community health
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    • v.16 no.1
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    • pp.61-69
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    • 1991
  • Serodiagnosis of Clonorchis sinensis infections will probably be a first choice tool for screening of clonorchiasis in a future because of increasing difficulties in collection and examination of stools. The sensitive test such as ELISA can he used effectively. However there are some limitations in serological diagnosis for the detection of serum antibody. One of the major problems is the non-specificity of the antigens which produce cross reaction with other helminthic infection sera. To solve this problem. many investigators have tried to purify the antigens used. In this study, we determined the antigenic profile of the crude saline extract antigen of C. sinensis at early developmental stage based on SDS-PAGE and immunoblotting techniques for the purpose of understanding the nature of C. sinensis worm antigen The following results were obtained : 1) The SDS-PAGE showed many protein hands ranging from 10Kd to 91Kd relative molecular weight. Among them, 66, 46, 40, 33, 27, 24, 16, 14 and 10Kd bands were observed as a principle bands. The protein components of C. sinensis changed chronologically during their early developmental period. 44Kd band was stained unclearly in antigen of 2 weeks worm, but changed to concentrated state in antigen of 5 weeks worm. 35Kd band was found in antigen of 2 weeks worm, however this band was disappeared in antigen of 5 weeks worm. 22Kd band also lost its staining property gradually. 2) In spite of differences in antigenic profile, there was no differences in the data obtained by microplate ELISA using each antigen preparation. Absorbance value began to rise in between 2 to 3 weeks after infection. 3) By EITB. serum antibody recognized major protein bands with molecular weight of 91, 85, 63, 46, 40, 33, 24, 14 and 10Kd hand respectively. Among them 66, 33, 17 and 14Kd bands were observed as non-specific band because they reacted even in normal control sera. Generally, gradual increase of positive reactions were observed as the infection period of C. sinensis was prolonged. In other hand, the reaction of 10Kd hand did not occurred when 26th week sera was tested. 4) The positive reactions using antigens of 2 weeks worm, especially on 40 and 24Kd bands, were most strong and sharply demarcated compared to those of 3~5 weeks worm antigen.

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Case of Acute Hemolytic Transfusion Reaction due to Anti-Fya Alloantibody in a Patient with Autoimmune Hemolytic Anemia (자가면역용혈환자에서 항-Fya 동종항체에 의한 급성용혈성수혈반응 1예)

  • Choi, Seung Jun;Nah, Hyunjin;Kim, Yundeok;Kim, Sinyoung;Kim, Hyun Ok
    • The Korean Journal of Blood Transfusion
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    • v.29 no.3
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    • pp.320-327
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    • 2018
  • A 72-year-old man with general weakness visited the outpatient clinic of the hematology department. The patient had been treated under the diagnosis of autoimmune hemolytic anemia for 2 years. His hemoglobin level at the time of the visit was 6.3 g/dL, and a blood transfusion was requested to treat his anemia. The patient's blood type was A, RhD positive. Antibody screening and identification test showed agglutination in all reagent cells with a positive reaction to autologous red blood cells (RBCs). He had a prior transfusion history with three least incompatible RBCs. The patient returned home after receiving one unit of leukoreduced filtered RBC, which was the least incompatible blood in the crossmatching test. After approximately five hours, however, fever, chills, dyspnea, abdominal pain, and hematuria appeared and the patient returned to the emergency room next day after the transfusion. The $anti-Fy^a$ antibody, which was masked by the autoantibody, was identified after autoadsorption using polyethylene glycol. He was diagnosed with an acute hemolytic transfusion reaction due to $anti-Fy^a$ that had not been detected before the transfusion. In this setting, it is necessary to consider the identification of coexisting alloantibodies in patients with autoantibodies and to become more familiar with the method of autoantibody adsorption.

Expression of Translationally Controlled Tumor Protein (TCTP) Gene of Dirofilaria immitis Guided by Transcriptomic Screening

  • Fu, Yan;Lan, Jingchao;Wu, Xuhang;Yang, Deying;Zhang, Zhihe;Nie, Huaming;Hou, Rong;Zhang, Runhui;Zheng, Wanpeng;Xie, Yue;Yan, Ning;Yang, Zhi;Wang, Chengdong;Luo, Li;Liu, Li;Gu, Xiaobin;Wang, Shuxian;Peng, Xuerong;Yang, Guangyou
    • Parasites, Hosts and Diseases
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    • v.52 no.1
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    • pp.21-26
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    • 2014
  • Dirofilaria immitis (heartworm) infections affect domestic dogs, cats, and various wild mammals with increasing incidence in temperate and tropical areas. More sensitive antibody detection methodologies are required to diagnose asymptomatic dirofilariasis with low worm burdens. Applying current transcriptomic technologies would be useful to discover potential diagnostic markers for D. immitis infection. A filarial homologue of the mammalian translationally controlled tumor protein (TCTP) was initially identified by screening the assembled transcriptome of D. immitis (DiTCTP). A BLAST analysis suggested that the DiTCTP gene shared the highest similarity with TCTP from Loa loa at protein level (97%). A histidine-tagged recombinant DiTCTP protein (rDiTCTP) of 40 kDa expressed in Escherichia coli BL21 (DE3) showed immunoreactivity with serum from a dog experimentally infected with heartworms. Localization studies illustrated the ubiquitous presence of rDiTCTP protein in the lateral hypodermal chords, dorsal hypodermal chord, muscle, intestine, and uterus in female adult worms. Further studies on D. immitis-derived TCTP are warranted to assess whether this filarial protein could be used for a diagnostic purpose.

Comparative Analysis of Screening Results from Various ELISA Formats Used for Detection of Anti-Erythropoietin Antibodies in Korean Patients

  • Ha, Sung-Kyu;Yang, Seung-Ju;Shin, Sug-Kyun;Jo, Young-Il;Baek, Kyung-Min;Hong, Seung-Hwa;Pack, Seung-Pil;Kim, Sung-Jo;Heo, Tae-Hwe
    • Biomolecules & Therapeutics
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    • v.18 no.2
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    • pp.184-190
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    • 2010
  • Clinical cases of pure red cell aplasia (PRCA) have been reported during the recombinant human erythropoietin (EPO) therapy for the anemia patients. PRCA is a rare hematological disorder leading to a severe anemia due to an almost complete stop of red blood cell production. Antibody (Ab)-associated PRCA is caused by the EPO-neutralizing Abs that eliminate the biological activity of EPO. In order to detect anti-EPO Abs in human sera, we performed conventional ELISA, directly coated bridging ELISA, and streptavidin coated bridging ELISA, and compared their sensitivity and specificity. Some false positive results were obtained in the conventional ELISA. One positive sample was detected successfully by streptavidin coated bridging ELISA, which was not appeared in the directly coated bridging ELISA. In conclusion, streptavidin coated bridging ELISA was substantially sensitive and specific format and one out of sixty-eight serum samples was proved to be anti-EPO positive.

Investigation of Immune Biomarkers Using Subcutaneous Model of M. tuberculosis Infection in BALB/c Mice: A Preliminary Report

  • Husain, Aliabbas A.;Daginawala, Hatim F.;Warke, Shubangi R.;Kalorey, Devanand R.;Kurkure, Nitin V.;Purohit, Hemant J.;Taori, Girdhar M.;Kashyap, Rajpal S.
    • IMMUNE NETWORK
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    • v.15 no.2
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    • pp.83-90
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    • 2015
  • Evaluation and screening of vaccines against tuberculosis depends on development of proper cost effective disease models along with identification of different immune markers that can be used as surrogate endpoints of protection in preclinical and clinical studies. The objective of the present study was therefore evaluation of subcutaneous model of M.tuberculosis infection along with investigation of different immune biomarkers of tuberculosis infection in BALB/c mice. Groups of mice were infected subcutaneously with two different doses : high ($2{\times}10^6CFU$) and low doses ($2{\times}10^2CFU$) of M.tuberculosis and immune markers including humoral and cellular markers were evaluated 30 days post M.tuberculosis infections. Based on results, we found that high dose of subcutaneous infection produced chronic disease with significant (p<0.001) production of immune markers of infection like $IFN{\gamma}$, heat shock antigens (65, 71) and antibody titres against panel of M.tuberculosis antigens (ESAT-6, CFP-10, Ag85B, 45kDa, GroES, Hsp-16) all of which correlated with high bacterial burden in lungs and spleen. To conclude high dose of subcutaneous infection produces chronic TB infection in mice and can be used as convenient alternative to aerosol models in resource limited settings. Moreover assessment of immune markers namely mycobacterial antigens and antibodies can provide us valuable insights on modulation of immune response post infection. However further investigations along with optimization of study protocols are needed to justify the outcome of present study and establish such markers as surrogate endpoints of vaccine protection in preclinical and clinical studies in future.

Immunohistochemical Detection of p53 Gene Mutation in Urine Samples in the Patients with Bladder Cancer (방광암 환자의 요세포 검사에서 p53 단백 발현의 의의)

  • Lee, Sang-Sook;Bae, Ji-Yeon;Kang, Yu-Na;Cho, Young-Rok;Park, Nam-Jo;Kim, Seun-Young;Kim, Jung-Hi
    • The Korean Journal of Cytopathology
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    • v.7 no.2
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    • pp.144-150
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    • 1996
  • Although bladder cancers are very common, little is known about their molecular pathogenesis. It is known that p53 alteration is found in about 60% of muscle-invasive bladder cancer, necessiating aggressive therapy and poor outcome. We examined the nuclear expression of p53 protein, using D07 monoclonal antibody in the urine samples from 31 patients with transitional cell carcinoma of the bladder to investigate the correlation of p53 overexpression with histologic grades and depth of invasion. The positive rate of p53 protein was 27% in superficial bladder tumor, but increased up to 71% in the invasive bladder carcinomas. The overexpression of p53 protein increased according to Mostofi grading system from 18% in grade I, 45% in grade II, and up to 100% in grade III. The p53 expression tended to be higher in the invasive and high grade bladder cancers than in the superficial and low grade ones(p<0.05). These results suggest that immunohistochemical analysis of the urine specimen in the bladder cancer patients could be a useful method of screening for the presence of p53 mutant protein. The mutant p53 protein expression may be an indicator of bladder cancer with more proliferative potential and/or aggressive biologic behavior.

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Recent Development of Protein Microarray and Proteogen Platform

  • Han, Moon-Hi;Kang, In-Cheol;Lee, Yoon-Suk;Cho, Yong-Wan;Lee, Eun-Kyoung
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.47-47
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    • 2005
  • There are many different surface technologies currently applied for preparation of protein chips. However, it requires innovative surface chemistry for capture proteins to be immobilized on chip surface keeping their conformation and activity intact and their orientation right, while they bind tightly and densely in a given array spot. Proteogen has developed 'ProteoChip BP' coated with novel proprietary linker molecules $(ProLinker^{TM})$ for efficient and robust immobilizations of capture proteins by improving surface properties of molecular captures. It was demonstrated that $ProLinker^{TM}$ gave the best surface performance in preparation of protein microarray chip base plates among others currently available on the market. In particular, the $ProLinker^{TM}-based$ surface chemistry has demonstrated to provide excellent performance in preparation of 'Antibody Chip' for analysis of biomarkers as well as proteome expression profiles. The linker molecule has also shown to be well applicable for development of biosensors and micro-beads as well as protein microarray and nano-array. ProteoChip BP can be used either for preparation of high-density array by using a microarrayer or for preparation of 'Well-on-a-Chip' with low density array, which is better applicable for quantitative analysis of biomarkers or protein-protein interactions. The biomarker assay can be performed either by direct or sandwich methods of fluorescence immunoassay. Application of ProteoChip BP has been well demonstrated by the extensive studies of 1) tumor-marker assays, 2) new drug screening by using 'Integrin Chip' and 3) protein expression profile analysis. Some of experimental results will be presented.

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Structure and Function of NtCDPK1, a Calcium-dependent Protein Kinase in Tobccco

  • Yoon, Gyeong-Mee;Lee, Sang-Sook;Pai, Hyun-Sook
    • Journal of Plant Biotechnology
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    • v.2 no.2
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    • pp.79-82
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    • 2000
  • We have isolated a cDNA encoding a calcium-dependent protein kinase (CDPK) in Nicotiana tabacum, which was designated NtCDPK1. Accumulation of the NtCDPK1 mRNA was stimulated by various stimuli, including phytohormones, CaCl$_2$ wounding, fungal elicitors, chitin and methyl jasmonate. The NtCDPK1 gene encodes a functional Ser/Thr protein kinase of which phosphorylation activity is strongly induced by calcium. By analyzing expression of the NtCDPK1-GFP fusion protein and by immunoblotting with antibody which reacts with NtCDPK1, we found that NtCDPK1 is localized in membrane and nucleus in plant cells. Silencing expression of the NtCDPK1 transgene resulted in marked decrease of lateral root development in the transgenic tobacco plants. Yeast two hybrid screening using NtCDPK1 as a bait identified a tobacco homologue of proteasome regulatory subunit 21D7, designated Nt21D7. The 21D7 mRNA has been shown to be predominantly expressed in proliferating tissues in the cell cycledependent manner in carrot. The recombinant NtCDPK1 protein associated with Nt21D7 in vitro, and could phosphorylate the Nt21D7 protein in vitro in the presence of calcium, suggesting that Nt21D7 protein is a natural substrate of NtCDPK1 in tobacco. These results suggest that NtCDPK1 may regulate tell proliferation processes, such as lateral root formation, by regulating specificity and/or activity of proteasome-mediated protein degradation pathway.

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A Case of Lupus Nephritis with Positive Antiphospholipid Antibodies, Initially Detected Through Analysis of Urinary Mass Screening (학교신체검사에서 발견된 항인지질 항체 양성 낭창성 신염 1례)

  • Lee Taek-Jin;Choi Min Sook;Lee Young-Mock;Kim Ji-Hong;Kim Pyung-Kil;Jeong Hyeon-Joo
    • Childhood Kidney Diseases
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    • v.5 no.2
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    • pp.219-224
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    • 2001
  • Antiphospholipid syndrome is a thrombotic disorder characterized by the association of arterial and venous thrombosis with the antibodies directed toward phospholipids. The presence of these antibodies in systemic lupus erythematosus(SLE) has been shown to be related to several clinical and analytical alterations. We experienced one case of lupus nephritis with positive antiphospholipid antibodies in a 10-year-old girl whose chief complaint was persistent microscopic hematuria. We report this case with a brief review of related literatures. (J. Korean Soc Pediatr Nephrol 2001 ;5 : 219-24)

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