• Asian Pacific Journal of Cancer Prevention
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• 제16권12호
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• pp.5115-5118
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• 2015

Background: The $p16^{INK4a}$ is a protein that expressed in Liquid-based cervical cytology specimens and has been proved link to cervical cancer. The $p16^{INK4a}$ could be detection by piezoelectric immunosensor and the immobilization of the $p16^{INK4a}$ antibody influence the sensitivity of the piezoelectric immunosensor. Materials and Methods: $5{\mu}L$ mouse polyclonal antibody against $p16^{INK4a}$ was bound onto the surface of immonosensor through two methods. (directly immobilized method; protein A method). Absorb of the $p16^{INK4a}$ antibody on the surface of immonosensor caused a shift in the resonant frequency of the immunosensor and The frequency changes recorded showed a better reproducibility. The activity of the immobilization antibody with the directly method and protein A method was tested with $p16^{INK4a}$ antigen. Results: The resonant frequency for different antibody immobilization methods were different, and the sensitivity for $p16^{INK4a}$ detection also different. Conclusions: The protein A method was found to be much more better than the directly method for the immobilization of the p16INK4A antibody on the gold electrode of the quartz crystal for cervical lesion detection. The Protein A method created more reproducible and stable immobilization antibody layers with p16INK4A antigen.

• Biotechnology and Bioprocess Engineering:BBE
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• 제8권4호
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• pp.227-232
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• 2003

A self-assembled monolayer of protein G was fabricated to develop an immunosensor based on surface plasmon resonance (SPR), thereby improving the performance of the antibodybased biosensor through immobilizing the antibody molecules (lgG). As such, 11-mercaptoundecanoic acid (11-MUA) was adsorbed on a gold (Au) support, while the non-reactive hydrophilic surface was changed through substituting the carboxylic acid group (-COOH) in the 11-MUA molecule using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrocholide (EDAC). The formation of the self-assembled protein G layer on the Au substrate and binding of the antibody and antigen were investigated using SPR spectroscopy, while the surface topographies of the fabricated thin films were analyzed using atomic force microscopy (AFM). A fabricated monoclonal antibody (Mab) layer was applied for detecting E. coli O157:H7. As a result, a linear relationship was achieved between the pathogen concentration and the SPR angle shift, plus the detection limit was enhanced up to 10$^2$ CFU/mL.

• BMB Reports
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• 제39권4호
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• pp.412-417
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• 2006

We examined the intracellular localization of NS5 protein of Dendrolimus punctatus cytoplasmic polyhedrosis virus (DpCPV) by expressing NS5-GFP fusion protein and proteins from deletion mutants of NS5 in baculovirus recombinant infected insect Spodoptera frugiperda (Sf-9) cells. It was found that the NS5 protein was present at the plasma membrane of the cells, and that the N-terminal portion of the protein played a key role in the localization. A transmembrane region was identified to be present in the N-terminal portion of the protein, and the detailed transmembrane domain (SQIHMVWVKSGLVFF, 57-71aa) of N-terminal portion of NS5 was further determined, which was accorded with the predicted results, these findings suggested that NS5 might have an important function in viral life cycle.

• Animal cells and systems
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• 제3권2호
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• pp.173-180
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• 1999

A blue biliprotein, insecticyanin (INS), has been purified from the last instar larval hemolymph of Agrius convolvuli by ultracentrifugation, Sephadex G-100 gel permeation chromatography, and preparative electrophoresis. The molecular mass of INS was estimated to be 26 kDa and the N-terminal sequence of INS revealed high similarity to that of Manduca sexta. Results of Western blotting and autoradiography indicated that INS is synthesized by the epidermis and released into the hemolymph. In contrast to the INS reported in other insects, Agrius convolvuli INS contained a small amount of lipid, predominately consisting of triacylglycerol. Subcellular localization of INS was determined using protein-A gold particles linked to secondary antibodies (anti-rabbit Ig). INS was heavily accumulated in the cytoplasmic inclusion body (CIB). CIBs showed a variety of shapes from rod to globule and generally surrounded the nucleus. They were mostly located near the basement membrane and especially abundant in the intersegmental membrane.

• 한국동물학회지
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• 제35권2호
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• pp.149-160
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• 1992

Using the protein A-gold complex, the mvoabrillogenesis and actin localization of cultured myoblast were invastisated. In the superstructural changes of mvogenic cell during differentiation, pectoral myoblasts contained large nucleus and numerous ribosomes but no myofibrils during the first 24 hr of cultures. Mvoblast initiated to differentiate at 3-day of culture contained the primitive myofibrillar structure. At 96 hr of culture, the mvofibrillar structure showed reletively discernable Z band but pools defined A, H and M bands. The feature of sarcomeric structure showed more defined form at cultur 5 day. In the aspect of actin localization, actin wvas diffusely detected throughout the cytoplasm of myogenic cell and nucleus during the proliferating stage. At 72 hr of culture, with the appearantc oi primitive mvofibrils, gold particles were observed in surrounding of myofibrils but still presented in overall of cytoplasm, especially in the surface and lumen of endoplasmic reticulum. With the gradual increase of culture time, local distribution of actin was readily detected within cytoplasm. In the 5-day specimen of cultures, gold particles precisely indicate the sites of actin localifation within the sarcomere. These results indicate the time of onset of myofibrill appearance and the biosynthetic and incorporation pathway of actin molecules into sarcomeric structure during myofibrillogenesis. Thus, in the present study, the first mvoabrillar structure was detected at culture 3 day, and the initiation of assembly into a typical sarcmeric structure was observed at culture 5 day. It seems, however, that the course of events on myofibrillogenesis of cultured myoblasts can be changed with great dependence of culture conditions including the number and groluth rate of mononucleated mvoblasts after seeding although the fundamental process shows identical appearances.

• Molecules and Cells
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• 제20권1호
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• pp.119-126
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• 2005

${\alpha}$-Expansins are bound to the cell wall of plants and can be solubilized with an extraction buffer containing 1 M NaCl. Localization of ${\alpha}$-expansins in the cell wall was confirmed by immunogold labeling and electron microscopy. The subcellular localization of vegetative ${\beta}$-expansins has not yet been studied. Using antibodies specific for OsEXPB3, a vegetative ${\beta}$-expansin of rice (Oryza sativa L.), we found that OsEXPB3 is tightly bound to the cell wall and, unlike ${\alpha}$-expansins, cannot be solubilized with extraction buffer containing 1 M NaCl. OsEXPB3 protein could only be extracted with buffer containing SDS. The subcellular localization of the OsEXPB3 protein was confirmed by immunogold labeling and electron microscopy. Gold particles were mainly distributed over the primary cell walls. Immunohistochemistry showed that OsEXPB3 is present in all regions of the coleoptile and root tissues tested.

• Tuberculosis and Respiratory Diseases
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• 제72권5호
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• pp.416-425
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• 2012

Background: The sensitivities and specificities of interferon-gamma release assays (IGRAs) vary among different population studies, and the data on the routine use of IGRAs are limited. The aim of this study was to evaluate the role of QuantiFERON-TB Gold In-Tube (QFT-GIT) test in the diagnosis of active tuberculosis. Methods: We conducted a prospective study, enrolling 77 patients with suspected pulmonary tuberculosis (TB), at a secondary care teaching hospital in Seoul. Results: In total, 12 (15.6%) patients showed indeterminate results due to positive control failure on the QFT-GIT test. Indeterminate results were significantly associated with the elderly, history of the intensive care unit stay, lymphocytopenia, especially low CD4 count, increased C-reactive protein and decreased protein levels. Of the 77 patients, 44 (57.1%) were diagnosed with active pulmonary tuberculosis, and the percentage of false negative results of the QFT-GIT was 36.4% (vs. 31.8% with TST). In the TB group with >65 years old (n=12), the proportions of the indeterminate (33.3% vs. 3.1%) and the false negative results (58.3% vs. 25.0%) of the QFT-GIT were significantly higher than in the younger TB group (n=32). Conclusion: Indeterminate and false negative results of QFT-GIT test were not infrequent in tuberculosis, especially in the elderly. Care should be considered for the interpretation with the elderly, immunocompromised, chronic and severely diseased patients.

• Applied Microscopy
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• 제28권4호
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• pp.527-538
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• 1998

This study was performed to investigate the activity of antibodies in the tissues of Paragonimus westermani at the different developmental stages. Enzyme-linked immunosorbent assay (ELISA) and Immunelectron microscopy (IEM) were applied, using the dog sera infected with metacercariae isolated from Cambaroides similis. These dog sera were obtained from 3rd to 96th week after infection by bleeding. The supernatants of homogenated worms for worm antigen were used. The worm tissues were embedded in Lowicryl HM 20 medium, treated with infected serum and protein A gold complex (particle size; 12 nm) and observed by electron microscope. In the pattern of antibody levels by ELISA test in all developemental worm antigens, the activity of antibody was very weak in the 3rd week, but strong in the worm antibody from 4th to 20th week after infection. Its activity was maintained even till 96th week. The antibody level of the L2th week worm antigen was higher than those of the 20th and 48th week worm antigens. Generally, many gold particles were observed on the secretory granules and the epithelial lamellae. Thus, it was concluded that the antigenic materials in the developmental worm tissues were especially concentrated on the secretory granules in the parenchymal tissues and the epithelial lamellae in the lumen of the caecum.

• Parasites, Hosts and Diseases
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• 제29권1호
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• pp.31-42
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• 1991

폐흡충(Paragonimus westermani)에 감염된 숙주에 대해 항체 생성을 유도하는 물질이 분포하고 있는 부위를 화인하기 위하여 대조군의 IgG와 폐홉충에 감염된 실험 개의 특이 IgG를 폐흡충 유약성충 조직에 반응시키고 면역황금표식법을 이용하여 전자현미경으로 관찰하여 다음과 같은 결과를 얻었다. 대조군의 IgG를 반응시킨 유약성충 표피층은 표피세포에서 분비되는 물질이 포함된 표피 합포체로 구성되어 기저층과 분명하게 구별되었으며 근육층도 잘 발달되어 있었다. 난황세포는 크기가 다양한 분비과립 등을 포함하고 있었으며 특히 조면소포체가 발달되어 있었다. 맹관의 막구조물은 잘 발달되었으며 장상피 합포체도 잘 발달되어 있었다. 한펀, 폐흡충에 감염된 실험동물의 특이 항체로 반응시킨 유약성충 표피충의 표피 합포체와 표피세포의 세포질에 황금입자의 매우 특이적인 표식가 관찰되었고 난황세포에서는 분필과립에 황금 입자가 높은 밀도로 관찰되었다. 그리고 맹관 상피 합포체는 미약하게 황금 표지가 되었고 맹관 막구조물과 맹관 내강에도 특이하게 황금 표지가 관찰되었지만 표피 합포체와 표피세포보다 밀도가 낮게 관찰되었다. 이상의 결과로 폐홉충에 감염된 실험 개는 폐흡충의 표피 합포체와 표피세포에서 생성된 물질과 난황세포에서 생성된 물질에 의하여 면역항체가 형성되며 일부는 맹관 내용물에 의해 약간의 면역항체가 유도되는 것으로 생각되었다.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2013년도 제45회 하계 정기학술대회 초록집
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• pp.251.2-251.2
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• 2013

Natural photosynthesis utilizes two proteins, photosystem I and photosystem II, to efficiently oxidize water and reduce NADP+ to NADPH. Artificial photosynthesis which mimics this process achieve water splitting through a two-step Z-schematic water splitting process using man-made synthetic materials for hydrogen fuel production. In this study, Z-scheme system was achieved from the hybrid materials which composed of hydrogen production part as photosystem I protein and water oxidizing part as semiconductor BiVO4. Utilizing photosystem I as the hydrogen evolving part overcomes the problems of existing hydrogen evolving p-type semiconductors such as water instability, expensive cost, few available choices and poor red light (>600 nm) absorbance. Some problems of photosystem II, oxygen evolving part of natural photosynthesis, such as demanding isolation process and D1 photo-damage can also be solved by utilizing BiVO4 as the oxygen evolving part. Preceding research has not suggested any protein-inorganic-hybrid Z-scheme composed of both materials from natural photosynthesis and artificial photosynthesis. In this study, to realize this Z-schematic electron transfer, diffusion step of electron carrier, which usually degrades natural photosynthesis efficiency, was eliminated. Instead, BiVO4 and Pt-photosystem I were all linked together by the mediator gold. Synthesized all-solid-state hybrid materials show enhanced hydrogen evolution ability directly from water when illuminated with visible light.