• Title/Summary/Keyword: monoclonal

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Application of Monoclonal Antibody to Develop Diagnostic Techniques for Infectious Bovine Rhinotracheitis Virus I. Production of Monoclonal Antibodies against Infectious Bovine Rhinotracheitis Virus (단(單)클론성 항체(抗體)를 이용한 소전염성비기관염(傳染性鼻氣管炎)바이러스 진단법(診斷法) 개발 I. 소전염성비기관염(傳染性鼻氣管炎)바이러스에 대한 단(單)클론성 항체(抗體) 생산(生産))

  • Jun, Moo Hyung;Kim, Duck Hwan;Lee, Hun Jun;An, Soo Hwan;Kweon, Chang Hee
    • Korean Journal of Agricultural Science
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    • v.14 no.2
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    • pp.401-408
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    • 1987
  • Nine monoclonal antibodies directed against infectious bovine rhinotracheitis virus (IBRV) were prepared by using cell hybridization technique, and the biological properties of the antibodies were investigated by means of immunofluorescence, serum neutralization, and electrophoretic analysis. Eight of 9 monoclonal antibodies reacted specifically with the antigenic constituents of IBRV, infectious laryngotracheitis virus, Marek's disease virus, turkey herpesvirus, hog cholera virus, porcine parvovirus and transmissible gastroenteritis virus. However, the remaining one, 26-2 clone, was found to be cross-reactive with pseudorabies virus. Two monoclonal antibodies, 7-C-2 and 12-A-2, which had neutralizing activity, were reactive with the molecular weights of 72 kilo daltons (72K) and 125K of IBRV proteins electrophoretically separated, respectively. The monoclonal antibody, 3-H-3, which is corresponding to 94K of IBRV proteins, revealed no neutralizing activity. The cross-reactive monoclonal antibody, 26-2, was proved by electrophoretical analysis to be reactive with 100K of IBRV proteins and 40K of pseudorabies virus.

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The Immunotyping Distribution of Serum Monoclonal Paraprotein and Environmental Impact on Multiple Myeloma (MM) and Monoclonal Gammopathy of Uncertain Significance (MGUS) in Taiwan: A Medical Center-Based Experience

  • Chang, Chih-Chun;Su, Ming-Jang;Lee, Shu-Jene;Tsai, Yu-Hui;Kuo, Lin-Yin;Lin, I-Hsin;Huang, Hui-Ling;Yen, Tzung-Hai;Chu, Fang-Yeh
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.1
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    • pp.395-399
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    • 2016
  • Background: Whether ambient exposure to environmental pollutants leads to hematopoietic malignancies such as multiple myeloma (MM) remains to be ascertained. Therefore, we aimed to investigate the immunotyping distribution of serum monoclonal paraprotein and the environmental influence on MM and monoclonal gammopathy of uncertain significance (MGUS) in the Taiwanese population. Materials and Methods: Serum protein electrophoresis with immunosubtraction by the capillary zone electrophoresis method was performed as primary screening for MM and MGUS. Clinical, pathological, and residence data of patients were also obtained. Results: From August, 2013 to June, 2015, a total of 327 patients underwent serum protein electrophoresis with immunosubtraction. Among these, 281 demonstrated no remarkable findings or non-malignant oligoclonal gammopathy, 23 were detected to have MGUS, 18 were identified as MM, and a further 5 were found as other malignancies. The most frequent immunotyping distribution of serum monoclonal paraprotein was IgG kappa (54.3%, n=25), followed by IgA lambda (15.2%, n=7) and IgG lambda (10.9%, n=5) in subjects with gammopathy. Additionally, it was shown that the elderly (OR: 4.61, 95% CI: 1.88-11.30, P<0.01) and males (OR: 2.04, 95% CI: 1.04-4.02, P=0.04) had significantly higher risk of developing MM and MGUS. There was no obvious impact of environmental factors on the health risk of MM and MGUS evolution (OR: 0.77, 95% CI: 0.40-1.50, P=0.49). Conclusions: The most frequent immunotyping distribution of serum monoclonal paraprotein included IgG kappa, IgA lambda and IgG lambda in MM and MGUS in the Taiwanese population. The elderly and male subjects are at significantly higher risk of MM and MGUS development, but there was no obvious impact of environmental factors on risk.

Purification of IgG1 Type Mouse Monoclonal Antibodies with DEAE-Trisacryl Chromatography (DEAE-Trisacryl 크로마토그래피법에 의한 IgG1 Type 쥐 단일클론 항체의 분리정제)

  • 최태부;정용근
    • Microbiology and Biotechnology Letters
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    • v.16 no.5
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    • pp.335-342
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    • 1988
  • An anion exchange chromatography was employed for the purification of mouse monoclonal antibodies from ascitic fluid and in vitro cultivation media. After cultivation of hybridomas, Alps 25-3, HCGK, A4W, and KW, producing IgG1, the culture supernatants were harvested by centrifugation, precipitated with 50-60% ammonium sulfate, and dialyzed against 0.025 M Tris-HCI buffer (pH 8.2). Then the dialyzed samples were loaded into a DEAE-Trisacryl M anion exchange column. Monoclonal antibodies bound to the DEAE-Trisacryl M were eluted with 0.025 M Tris-HCI buffer (pH 8.2) containing 30-40 mM NaCl. In ammonium sulfate precipitation, the recovery of the monoclonal antibody was shown to be 90% and 84% from in vitro culture media containing 10% and 2% fetal bovine serum, respectively. On the other hand, the pretreatment by ultrafiltration enhanced the yield up to 91% whereas the purity was lower than that by ammonium sulfate treatment. Subsequently, in the DEAE-Trisacryl M chromatographic separation, the purities and recoveries of all the monoclonal antibodies from both the in vitro culture supernatants and ascitic fluids were 70-80% and 65% respectively. The monoclonal antibody, Alps 25-3 could be further purified with a purity of 95% through an immunoadsorbent chromatography.

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Production and Evaluation of Monoclonal Antibodies Against Recombinant Coat Protein of Lily mottle virus for Western Blotting and Immono-blot Analysis

  • Chung, Bong-Nam;Yoon, Ju-Yeon;Choi, Gug-Sun
    • The Plant Pathology Journal
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    • v.25 no.3
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    • pp.225-230
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    • 2009
  • Lily mottle virus (LMoV) causes flower quality reduction in Lilium spp. The coat protein gene was RT-PCR-amplified from total RNA extracted from infected lily leaves and the amplified fragment was cloned into the pRSET expression vector tagged with a His-MBP. The plasmid of recombinant coat protein was used to transform an Escherichia coli strain pLysS and was expressed. The coat protein was purified by affinity chromatography using a Ni-NTA resin. The identity of the purified protein was confirmed by SDS-PAGE. The in vitro-expressed protein was used for immunization of mice. The polyclonal and monoclonal antibodies reacted specifically for the detection of LMoV in lily extracts in Western blot. Moreover the monoclonal antibodies reacted with lily extracts in DAS-ELISA with no unspecific or heterologous reactions against other non-serologically related viruses, but the polyclonal antibodies revealed a weak reaction against both infected lily and healthy control.

Demonstration of TCM-9 Monoclonal Antibody in Follicular Neoplasm of Thyroid (갑상선의 여포상 종양의 감별에 있어서 TCM-9의 발현양상)

  • Kim, Yun-Jung;Shim, Jung-Weon;Ahn, Hye-Kyung;Park, Young-Euy
    • The Korean Journal of Cytopathology
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    • v.7 no.2
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    • pp.134-137
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    • 1996
  • Monoclonal antibody(TCM-9) against human thyroid cancers have been studied by screening with human thyroid cancers, normal and benign thyroid tissue, and normal human serum protein. A monoclonal antibody(TCM-9) that is known to have strong specificity for human thyroid cancer but not for Graves' disease, adenoma or normal thyroid does not bind to native or mature human thyroglobulin(Tg). We used to TCM-9 antibody by immunohistochemical staining on 5 follicular cancer, 2 follicular adenoma, 1 follicular neoplasm with suspicious invasion, 2 papillary cancer to ascertain being of help in differentiation between follicular carcinoma and adenoma. Reactivity of TCM-9 was observed in follicular carcinoma and papillary carcinoma but not observed in follicular adenoma. Thus TCM-9 is a novel monoclonal antibody against the thyroid cancer.

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Sensorimotor Polyneuropathy and Multiple Cranial Neuropathies Associated with IgG κMonoclonal Gammopathy of Undetermined Significance (IgG κMonoclonal Gammopathy of Undetermined Significance와 연관된 운동신경 다발성 신경병증과 다발선 뇌신경병증)

  • Jeon, Seong-Ho;Kim, Yoon-Bong;Lee, Yeong-Bae;Park, Jong-Ho;Shin, Won-Chul;Park, Hyeon-Mi;Shin, Dong-Jin
    • Annals of Clinical Neurophysiology
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    • v.6 no.1
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    • pp.48-51
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    • 2004
  • The occurrence of polyneuropathy in association with monoclonal gammopathy of undetermined significance (MGUS) is quite common. However, reports of MGUS associated cranial neuropathies are rare. A 63 year-old women was presented with diplopia and swallowing difficulty. Neurological examination showed limitation of abduction of right eye, right peripheral facial palsy, decreased hearing and gag reflex, left side deviation of uvula, and decreased DTR. Sensorimotor polyneuropathy were observed with elctrophysiological studies. Protein and immunoelectrophoresis revealed IgG ${\kappa}$monoclonal gammopathy. She was treated with intravenous immunoglobulin and steroid, and her symptoms and signs were improved. This case suggested that she had sensorimotor polyneuropathy and multiple cranial neuropathies associated with IgG ${\kappa}$MGUS.

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Production of Monoclonal Antibodies to Phytohormones and Its Application (식물홀몬에 대한 단클론성 항체 생산과 이용)

  • 황태익
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.34 no.s01
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    • pp.48-54
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    • 1989
  • An immunoassay techniques using monoclonal antibodies have been developed of the quantitative analysis of endogenous hormones in plants. In order to be useful for immunoassay, both a high degree of specificity and a high affinity are required. A system is described for production of hybridomas which secrete antibodies against the plant hormone. Using the system we were able to produce hybridmas with the desired antibody specificity by cell fusion and culture method. For a number of obvious reasons, monoclonal antibodies(mAb) were superior to polyclonal antibodies.

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Production of Monoclonal Antibodies by Hybridomas Sensitized to Sporozoites of Cryptosporidium parvum (Cryptosporidium parvum Sporozoites 에 감작된 Hybridomas 에서의 Monoclonal Antibody 생산)

  • Cho, Myung-Hwan
    • Microbiology and Biotechnology Letters
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    • v.17 no.5
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    • pp.494-498
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    • 1989
  • Hybridoma cell lines, which secrete monoclonal antibodies (mAbs) against the surface antigens of Cryptosporidium parvum Sporozoites, were produced by fusing spleen cells of C. parvum Sporozoite-immunized mice with P3-X63-Ag8 myeloma cells. Two cloned antibody-secreting cell lines, Kor1 and Ea2, were established and produced IgG1 and IgG2a antibodies, respectively. Percoll-purified sporozoites were solubilized and separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Western blot assay demonstrates that an antigen of 20-kDa was bound by monoclonals. By indirect immunofluorescence microscopy, mAb exhibited uniform binding to the sporozoite surface.

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Immunoradiometric Assay using Monoclonal Antibody Against Human Serum Transferrin Receptor for Diagnosis of Iron Deficiency (사람 혈청 트란스페린수용체의 단클론 항체를 이용한 방사면역측정과 철영양상태의 진단)

  • 김승렬
    • Journal of Nutrition and Health
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    • v.29 no.9
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    • pp.971-980
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    • 1996
  • The soluble transferrin receptor(TfR) in human serum has been shown recently to be a truncated form of intact membrane bound receptor containing most of the extracellular domain. We purfied the transferin-free TfR from human serum by immounoaffinity chromatography which produced the single protein identity in high resolution gel chormatography. The monoclonal antibodies(MAb) against purifed serum TfR were produced by fusion of spleen cells o fimmunized Balb/c mice and SP2 cells. Ten hybrids producing MAb specific for serum TfR were identifed and determine their iostypes. A immunoraddiometric assay (IRMA) for serum TfR was established using two monoclonal IgG1 antibodies as the coating and indicator antibodies on the bosis of their suitability in sandwich IRMA of serum TfR. The mean serum TfR levels in the 15 normal male, 15 normal female, and 19 iron-deficient subjects were 5.4$\pm$0.98, 4.6$\pm$0/76, and 18.0$\pm$12.8mg/1, respectively, and the difference in mean values between normal and iron deficient subjects was significant(p=0.0005). There existed the inverse logarithmic relationship(r=-0.9336, p<0.0001) between the serum TfR and ferritin levels.

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Fine Mutational Analysis of 2B8 and 3H7 Tag Epitopes with Corresponding Specific Monoclonal Antibodies

  • Kim, Tae-Lim;Cho, Man-Ho;Sangsawang, Kanidta;Bhoo, Seong Hee
    • Molecules and Cells
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    • v.39 no.6
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    • pp.460-467
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    • 2016
  • Bacteriophytochromes are phytochrome-like light-sensing photoreceptors that use biliverdin as a chromophore. To study the biochemical properties of the Deinococcus radiodurans bacteriophytochrome (DrBphP) protein, two anti-DrBphP mouse monoclonal antibodies (2B8 and 3H7) were generated. Their specific epitopes were identified in our previous report. We present here fine epitope mapping of these two antibodies by using truncation and substitution of original epitope sequences in order to identify minimized epitope peptides. The previously reported original epitope sequences for 2B8 and 3H7 were truncated from both sides. Our analysis showed that the minimal peptide sequence lengths for 2B8 and 3H7 antibodies were nine amino acids (RDPLPFFPP) and six amino acids (PGEIEE), respectively. We further characterized these peptides in order to investigate their reactivity after single deletion and single substitution of the original peptides. We found that single-substituted 2B8 epitope (RDPLPAFPP) and dual-substituted 3H7 epitope (PGEIAD) showed significantly increased reactivity. These two antibodies with high reactivity for the short modified peptide sequences are valueble for developing new peptide tags for protein research.