• Title/Summary/Keyword: human antibodies

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The Improved Antigen-binding Activity of Biosimilar Remicade ScFv Antibodies by Fusion of the Leucine Zipper Domain (Leucine zipper도메인의 융합에 의한 바이오시밀러 레미케이드 Single-chain Fv 항체의 항원 결합력 개선)

  • Kim, Jin-Kyoo;Kim, Tae Hwan
    • Journal of Life Science
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    • v.30 no.11
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    • pp.1012-1020
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    • 2020
  • Remicade is a therapeutic biosimilar natural antibody in which the mouse variable domain has been linked to the human constant domain. It is a chimeric monoclonal antibody specific to tumor necrosis factor-alpha (TNF-α) and has been developed for the treatment of rheumatoid arthritis. To investigate the biological activity of the Remicade antibody, we carried out a bioinformatics study using a protein data bank to characterize the TNF-α antigen binding mechanism of the Remicade natural antibody. Because the production of the Remicade antibody is often limited by genetic instability of the natural antibody-producing cell, we generated a Remicade single-chain variable domain fragment antibody (Remicade) in which a heavy chain variable domain (VH) is joined with a light chain variable domain (VL) by a polypeptide linker. Furthermore, Remicade was fused to a leucine zipper (RemicadeScZip) for higher production and higher antigen-binding activity than Remicade. The Remicade and Remicade ScZip were expressed in Escherichia coli and purified by a Ni+-NTA-agarose column. As expected, the purified proteins had migrated as 28.80 kDa and 33.96 kDa in sodium dodecyl sulfate-polyacrylamide electrophoresis. The TNF-α antigen binding activity of Remicade was not observed by ELISA and western blot. In contrast, RemicadeScZip showed antigen-binding activity. Additional bio-layer interferometry analysis confirmed the antigen-binding activity of RemicadeScZip, suggesting that the leucine zipper stabilized the folding of RemicadeScZip in a denatured condition and improved the TNF-α antigenbinding activity.

A Study on Needle Stick Injuries in Health Professionals (의료인의 바늘자상 실태에 관한 연구)

  • 김영분
    • Journal of Korean Academy of Nursing
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    • v.26 no.3
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    • pp.605-622
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    • 1996
  • Needle stick injury, in which blood-borne pathogens including Human Immune-Deficiency virus and hepatitis B virus are transmitted, is one of the major occupational hazards that health professionals face everyday. In order to provide basic data for the development of educational programs for health professionals aimed at preventing and effectively managing needle stick injuries, a retrospective descriptive study was carried out .The subjects of the study were 630 health professionals, 499 nurses and 131 physicians, from two university hospitals in Seoul, Korea. Data on episodes of needle stick experiences over the past is months September 1994 through August 1995, were collected between September 1 and 7, 1995. A Questionaire developed by the researcher was used. The frequencies and the percentile score for episodes of needle stick injuries were calculated using the PC-SAS program. The differences and similarities in reference to the structure, career, and specialty variables were analysed by X$^2$-tests. Results are as follows : 1. Of the sample, 521(82.7%) reported a needle stick injury, 33.4% reported 3 or more episodes of needle stick injuries. 2. The needle stick injuries occured in the following processes : process of percutaneous venepuncture for intra-venous injection and infusion(55.3%), medical examination and treatment(48.9%), per-cutaneous venepuncture for blood sampling (46.3%) and intra-muscular injection(42.2%). 3. The study showed that needle stick injuries occured before(19%), during(25%), and after (56%) client treatment. The major causes of needle stick injuries were perceived to be hastiness(82.2%) and carelessness(48.3%). Of these injuries, 91.8% occured in emergency situations. 4. Follow of care for the injury consisted of : treating the injured site immediately using disinfectants(89.7%), reviewing the clinical records of the patient involved(84.2%), immunological investigation for the status of antibodies(11.1%) and self-medication of antibiotics (10.7%). Only 16.3% of the total episodes were founded to have been reported to the administrative unit. 5. The length of clinical experience of the nurses, clinical specialty and length of clinical experience in physicians were found to have influenced the episodes of needle stick injuries ; nurses with less than 1 year and with more than 6 years of clinical experiences had significantly lower levels (X$^2$=25.04, P=.00), surgeons had significantly higher levels (X$^2$=9.89, P=.02) compared to that of internists and interns, higher(X$^2$=4.54, P=.03)than residents.

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The Effect of Gefitinib on Immune Response of Human Peripheral Blood Monocyte-Derived Dendritic Cells (인간 말초혈액 단핵구 유래 수지상세포의 면역반응에 미치는 Gefitinib의 영향)

  • Cho, Jin-Hoon;Kim, Mi-Hyun;Lee, Kwang-Ha;Kim, Ki-Uk;Jeon, Doo-Soo;Park, Hye-Kyung;Kim, Yun-Seong;Lee, Min-Ki;Park, Soon-Kew
    • Tuberculosis and Respiratory Diseases
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    • v.69 no.6
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    • pp.456-464
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    • 2010
  • Background: Synergistic antitumor effects of the combined chemoimmunotherapy based on dendritic cells have been reported recently. The aim of this study is to search new applicability of gefitinib into the combination treatment through the confirmation of gefitinib effects on the monocyte derived dendritic cells (moDCs); most potent antigen presenting cell (APC). Methods: Immature and mature monocyte-derived dendritic cell (im, mMoDC)s were generated from peripheral blood monocyte (PBMC) in Opti-MEM culture medium supplemented with IL-4, GM-CSF and cocktail, consisting of TNF-${\alpha}$ (10 ng/mL), IL-$1{\beta}$ (10 ng/mL), IL-6 (1,000 U/mL) and $PGE_2$ ($1{\mu}/mL$). Various concentrations of gefitinib also added on day 6 to see the influence on immature and mature MoDCs. Immunophenotyping of DCs under the gefitinib was performed by using monoclonal antibodies (CD14, CD80, CD83, CD86, HLA-ABC, HLA-DR). Supernatant IL-12 production and apoptosis of DCs was evaluated. And MLR assay with $[^3H]$-thymidine uptake assay was done. Results: Expression of CD83, MHC I were decreased in mMoDCs and MHC I was decreased in imMoDCs under gefitinib. IL-12 production from mMoDCs was decreased under $10{\mu}M$ of gefitinib sinificantly. Differences of T cell proliferation capacity were not observed in each concentration of geftinib. Conclusion: In spite of decreased expressions of some dendritic cell surface molecules and IL-12 production under $10{\mu}M$ of gefitinib, significant negative influences of gefitinib in antigen presenting capacity and T cell stimulation were not observed.

A Study on Rehabilitation Treatment Using Radiofrequency Treatment (고주파 통증치료기를 이용한 재활 치료에 대한 연구)

  • Jo, Jae-Hyun;Lee, Sang-Yong;Lee, Geun-Yong;Yoon, Se-Jin;Cheong, Ha-Young;Lee, Sang-Sik
    • The Journal of Korea Institute of Information, Electronics, and Communication Technology
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    • v.13 no.3
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    • pp.212-218
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    • 2020
  • When Radiofrequency energy is applied to the human body, the vibration width is very short. Therefore, the electrolyte burn generated when using the direct current does not occur. Ion molecules, polarized molecules, etc., vibrate more than 40,000 times per second, converting them into frictional heat to generate deep heat. The blood flow of capillaries increases 4-5 times more than at rest, increasing the supply of oxygen, nutrients, antibodies, and white blood cells. In addition, the electrochemical reaction does not occur because the vibration width and the pulsation period are very short. It is a physical factor treatment method that does not stimulate the sensory nerves and motor nerves. In this study, an isotonic exercise is performed in a young normal adult using a Radiofrequency pain treatment device. The purpose of this study is to integrate rehabilitation therapy by measuring electromyography data during isotonic exercise and confirming the effect on changes in motor neuron response. The EMG data generated when isotonic exercise of the forearm biceps muscle and the EMG data measured after the use of a Radiofrequency pain treatment device after exercise were RMS, respectively, and verified through t-test. It was confirmed that there was a significant difference in both men and women because the t-value was smaller than the significance level p (<.05).

Mucosal Immunity Related to FOXP3+ Regulatory T Cells, Th17 Cells and Cytokines in Pediatric Inflammatory Bowel Disease

  • Cho, Jinhee;Kim, Sorina;Yang, Da Hee;Lee, Juyeon;Park, Kyeong Won;Go, Junyong;Hyun, Chang-Lim;Jee, Youngheun;Kang, Ki Soo
    • Journal of Korean Medical Science
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    • v.33 no.52
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    • pp.336.1-336.12
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    • 2018
  • Background: We aimed to investigate mucosal immunity related to forkhead box P3 ($FOXP3^+$) regulatory T (Treg) cells, T helper 17 (Th17) cells and cytokines in pediatric inflammatory bowel disease (IBD). Methods: Mucosal tissues from terminal ileum and colon and serum samples were collected from twelve children with IBD and seven control children. Immunohistochemical staining was done using anti-human FOXP3 and anti-$ROR{\gamma}t$ antibodies. Serum levels of cytokines were analyzed using a multiplex assay covering interleukin $(IL)-1{\beta}$, IL-4, IL-6, IL-10, IL-17A/F, IL-21, IL-22, IL-23, IL-25, IL-31, IL-33, interferon $(IFN)-{\gamma}$, soluble CD40L, and tumor necrosis factor-${\alpha}$. Results: $FOXP3^+$ Treg cells in the lamina propria (LP) of terminal ileum of patients with Crohn's disease were significantly (P < 0.05) higher than those in the healthy controls. $ROR{\gamma}t^+$ T cells of terminal ileum tended to be higher in Crohn's disease than those in the control. In the multiplex assay, serum concentrations (pg/mL) of IL-4 ($9.6{\pm}1.5$ vs. $12.7{\pm}3.0$), IL-21 ($14.9{\pm}1.5$ vs. $26.4{\pm}9.1$), IL-33 ($14.3{\pm}0.9$ vs. $19.1{\pm}5.3$), and $IFN-{\gamma}$ ($15.2{\pm}5.9$ vs. $50.2{\pm}42.4$) were significantly lower in Crohn's disease than those in the control group. However, serum concentration of IL-6 ($119.1{\pm}79.6$ vs. $52.9{\pm}39.1$) was higher in Crohn's disease than that in the control. Serum concentrations of IL-17A ($64.2{\pm}17.2$ vs. $28.3{\pm}10.0$) and IL-22 ($37.5{\pm}8.8$ vs. $27.2{\pm}3.7$) were significantly higher in ulcerative colitis than those in Crohn's disease. Conclusion: Mucosal immunity analysis showed increased $FOXP3^+$ T reg cells in the LP with Crohn's disease while Th17 cell polarizing and signature cytokines were decreased in the serum samples of Crohn's disease but increased in ulcerative colitis.

Potential of Fucoidan Extracted from Seaweeds as an Adjuvant for Fish Vaccine (해조류 유래 Fucoidan의 어류용 백신 항원보조제로서의 가능성에 대한 고찰)

  • Min, Eun Young;Kim, Kwang Il;Cho, Mi Young;Jung, Sung-Hee;Han, Hyun-Ja
    • Journal of Marine Life Science
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    • v.4 no.1
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    • pp.1-13
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    • 2019
  • Fucoidan is a physiologically functional ingredient extracted from seaweed brown algae, which is a sulfated polysaccharide containing fucose as a main molecule backbone. Fucoidan has a variety of immune-modulating or -stimulating effects, including promoting antigen uptake and enhancing anti-bacterial, anti-viral and anti-tumor effects. In addition, recent studies have suggested the possibility of use of fucoidan as a vaccine adjuvant in the field of human vaccine. Use of fucoidan as supplementary feeds have already been studied, but the development of fucoidan as an adjuvant of fish vaccine is still premature. However, the intracellular uptake of fucoidan differs depending on the molecular weight of fucoidan, and there is a limit to the study on specific immune response including the production of antibodies to fish caused by an artificial infection of pathogen. Although the safety of fucoidan has been demonstrated in animal cells, there is a need to confirm the safety of fucoidan in fish. Therefore, active research in this field is needed to use fucoidan as a vaccine adjuvant. This study discussed the effects of fucoidan on immune stimulation, humoraland cellular- immunity including humans and animals. The prospect of fucoidan as a vaccine adjuvant in fisheries also reviewed.

Relationship between the Thyroid Hormone and Viral Infections in Pregnancy (임신 중 바이러스성 감염요인과 갑상선 호르몬의 상관성)

  • Lim, Dong-Kyu;Park, Chang-Eun
    • Korean Journal of Clinical Laboratory Science
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    • v.54 no.1
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    • pp.28-37
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    • 2022
  • Pregnancy requires an important interpretation of thyroid function tests. The presence of anti-thyroid antibodies and viral infectious agents affect the health of both the fetus and the mother. Hence, a selective evaluation of thyroid function in pregnancy is required. This study is a retrospective cross-sectional survey to examine the correlation between thyroid hormones and viral infections during pregnancy. The results showed that the triiodothyronine (T3) decreased with increasing age, especially in the hepatitis C virus (HCV)-positive group (P<0.01). In addition, although negative for the human immunodeficiency virus (HIV), thyroxine (FT4) showed a significant increase in near-threshold or twin pregnant women (P<0.05). The thyroid stimulating hormone (TSH) was highly distributed at the age of 30, and there was no statistically significant correlation with other viral infection factors. In addition, as a result of dividing and analyzing the result of TSH by the quantiles, FT4 and T3 showed a positive correlation but showed a negative correlation with TSH (P<0.05). Therefore, the evaluation of prenatal thyroid screening during pregnancy and viral infection factors should reflect the time of pregnancy, exposure to infection, and the quantitative values. Adequate thyroid hormone and viral infections availability is important for an uncomplicated pregnancy and optimal fetal development.

Production and Evaluation of Immunoreactivity of Poly Lysine-Tagged Single Chain Fragment Variable (ScFv) Lym-1 Antibody for Direct Conjugation to Fluorescence Dye (형광 물질 직접 표지를 위한 Poly Lysine 도입 Lym-1 단일사슬 항체의 제조 및 면역반응성 평가)

  • Jung, Jae-Ho;Choi, Tae-Hyun;Woo, Kwang-Sun;Chung, Wee-Sup;Kang, Joo-Hyun;Jeong, Su-Young;Choi, Chang-Woon;Lim, Sang-Moo;Cheon, Gi-Jeong
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.5
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    • pp.487-494
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    • 2009
  • Purpose: Small size of recombinant scFv antibody has many advantages such as rapid blood clearances and improved targeting antibodies to tumor region. On the other hand owing to small size, number of amino group is insufficient in conjugation with chelator and fluorescence labeling. This study is to introduce poly lysine tag to the C-terminal end of scFv lym-1 sequence for fluorescence chelator conjugation. Materials and Methods: Poly lysine scFv lym-1 gene, cloned into pET-22b (+) vector, was expressed in E. coli BL21 (DE3) strain. Antibody purification was performed with Ni-NTA column and then size exclusion column chromatography. Expression and purification levels of poly lysine tagged scFv lym-1 antibody were confirmed by western blot analysis. I-124, I-125, I-131 and Tc-99m were used for radiolabeling of purified poly lysine scFv lym-1. Flow cytometry analysis of FIT( conjugated poly lysine scFv lym-1 was performed for confirmation of immunoreactivity of human Burkitt's lymphoma cells. Results: Poly lysine scFv lym-1 antibody was purified through two steps and identified as molecular weight of 48 KDa. Radiolabeling yields of I-124, I-125, I-131 and Tc-99m into poly lysine scFv lym-1 were >99%, >99%, >95% and >99%, respectively. Flow cytometry analysis of poly lysine scFv and scFv lym-1 was showed similar immunoreactivity to human Burkitt's lymphoma cells. Conclusion: Poly lysine tag was useful for the sufficient number of amino groups to scFv lym-1 antibody for chelator conjugation with minimizing loss of immunoreactivity.

A MOLECULAR BIOLOGIC STUDY ON BIOCOMPATIBILITY OF METALLIC DENTAL MATERIALS USED FOR CHILDREN WITH CULTURED HUMAN GINGIVAL FIBROBLASTS (인체 섬유모세포(HGF-1) 배양에서 소아용 치과금속재의 세포친화성에 대한 분자생물학적 연구)

  • Kim, Ju-Mi;Jeong, Tae-Sung;Kim, Shin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.29 no.2
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    • pp.243-254
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    • 2002
  • For the purpose of evaluating the biocompatability of 3 kinds of metallic materials frequently used in pediatric dentistry (stainless steel crown, orthodontic band, orthodontic wire), cellular and molecular studies, including cell growth and proliferation, screening of cell death with determination of types whether necrosis or apoptosis and changes in expressions of related signaling molecules were examined, using cultured human gingival fibroblasts (HGF-1), HGF-1 was cultured in Dulbecco's modified Eagle's medium. among which the 3rd to 6th generations of HGF-1 were used. The specimen were divided into stainless steel crown (R), band (B) and wire (W). The immunocytochemical study was done for the detection of anti-PCNA (proliferating cell nuclear antigen) labeling. With extracted protein, western blot was done for the detection of ERK1/2, JNK, and p38, using individual antibodies. Cultured cells proliferated, remarkably till 7 day and slightly at 11 day. There was no statistical significance in the counts of proliferating HGF-1 between control and experimental groups (p>0.05). Relative growth rates were no statistically significant difference between control and experimental groups (p>0.05). PCNA labeling indexes showing similar patterns in control and experimental groups. The expressions of ERK1 and ERK2, p38 were similar in control and experimental groups. The expression of JNK increased at 1st day, slightly decreased at 4th day and markedly increased at 7th and 11 day. Although the patterns of control and experimental groups were similar, the increased expressions of JNK at late period suggest a possible stress due to inhibited cell growth and proliferation, and worse culture condition. Conclusively, the 3 kinds of metal specimens used in this study did not induce cellular and molecular hazards during short term culture of HGF-1. But, for the better clinical stability, the establishment of long period culture and animal experiment was thought necessary.

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Altered Expression of Peroxiredoxin and Thioredoxin in Septic Animal Model (패혈증 동물 모델에서 Peroxiredoxin 및 Thioredoxin의 발현 변화)

  • Kim, Hyung-Jung;Chae, Ho-Zoon;Ahn, Chul-Min;Kim, Sung-Kyu;Lee, Won-Young
    • Tuberculosis and Respiratory Diseases
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    • v.47 no.4
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    • pp.451-459
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    • 1999
  • Background : In sepsis, excessive generation of reactive oxygen species plays key roles in the pathogenesis of acute lung injury. The serum antioxidants such as catalase and MnSOD are elevated in sepsis and considered as predictors of acute respiratory distress syndrome(ARDS) and prognostic factors of sepsis. Peroxiredoxin(Prx) has recently been known as an unique and major intracellular antioxidant. In this study, we evaluated the expression of Prx I and Prx II in mouse monocyte-macrophage cells(RAW 267.7) after treatment of oxidative stress and endotoxin and measured the amount of Prx I, Prx II and thioredoxin(Trx) in peritoneal and bronchoalveolar lavage fluid of septic animal model. Methods : Using immunoblot analysis with specific antibodies against Prx I, Prx II and Trx, we evaluated the distribution of Prx I and Prx II in human neutrophil, alveolar macrophage and red blood cell. We evaluated the expression of Prx I and Prx II in mouse monocyte-macrophage cells after treatment of $5\;{\mu}M$ menadione and $1\;{\mu}g/ml$ lipopolysaccharide(LPS) and measured the amount of Prx I, Prx II and Trx in peritoneal lavage fluid of intraperitoneal septic animals(septic animal model induced with intraperitoneal 6 mg/Kg LPS injection) and those in bronchoalveolar lavage fluid of intraperitoneal septic animals and intravenous septic animals(septic animal model induced with intravenous 5 mg/Kg LPS injection) and compared with the severity of lung inflammation. Results : The distribution of Prx I and Prx II were so different among human neutrophil, alveolar macrophage and red blood cell. The expression of Prx I in mouse monocyte-macrophage cells was increased after treatment of $5\;{\mu}M$ menadione and $1\;{\mu}g/ml$ lipopolysaccharide but that of Prx II was not increased. The amount of Prx I, Prx II and Trx were increased in peritoneal lavage fluid of intraperitoneal septic animals but were not increased in bronchoalveolar lavage fluid of intraperitoneal and intravenous septic animals regardless of the severity of lung inflammation. Conclusion : As intracellular antioxidant, the expression of Prx I is increased in mouse monocyte-macrophage cells after treatment of oxidative stress and endotoxin. The amount of Prx I, Prx II and Trx are increased in local inflammatory site but not increased in injured lung of septic animal model.

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