• Journal of Microbiology and Biotechnology
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• v.16 no.1
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• pp.141-144
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• 2006

An antibody layer was fabricated to detect Escherichia coli O157:H7. The micropattern of 16-mercaptohexadecanoic acid (16-MHDA) as alkylthiolate was formed on the gold surface by using the PDMS stamp with microcontact printing $({\mu}CP)$ techniques. In order to form antibody patterns on the template, protein G was chemically bound to the 16-MHDA patterns, and antibody was adsorbed on a self-assembled protein G layer. The formation of the 16-MHDA micropattern, self-assembled protein G layer and antibody pattern on Au substrate was confirmed by surface plasmon resonance (SPR) spectroscopy. Finally, the micropatterning method was applied to fabricate the antibody probe for detection of E. coli O157:H7, and monitoring of antigen by using this probe was successfully achieved.

• Biomedical Science Letters
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• v.2 no.2
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• pp.257-265
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• 1996

Simple stain and electron microscopic in situ hybridization is studied and applied for the identification of rabbit haemorrhagic disease viral RNA in a unicrylated preparation of the liver after innoculation of rabbit haemorrhagic disease virus. Hybridization for detection of viral RNA in unicryl embedded tissues using complementary 84 bases oligonucleotide probe labelled by biotin CE-phosphoramidite compared with 4717∼4800 sequences of rabbit haemorrhagic disease virus, modified hybridization protocol and antibiotin antibody-l0nm gold as signal marker. The best results were obtained in 0.02% glutaraldehyde, Unicryl resin cell block, biotinylated oligonucleotide probes, antibiotin-l0nm gold. In this report, RHD viral RNA was distributed widely within the mitochondria and nucleus of liver cell by electron microscopic in situ hybridization. In situ hybridization has become a standard method for localizing DNA or RNA sequences in tissue or celt preparation. In situ hybridization is detected the virus genome in the cells and tissue as specifically compared with others nucleic acid hybridization method.

• Bulletin of the Korean Chemical Society
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• v.34 no.10
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• pp.2921-2924
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• 2013

Immunoliposomes (antibody-conjugated liposomes) are highly useful as both a drug carrier in drug delivery and as a reporting probe in immunodiagnostics. However, antibody conjugation is lengthy and cumbersome, because this includes several steps such as derivatization of the antibody, conjugation of the derivatized antibody to liposomes, and separation of the unbound antibodies from immunoliposomes. Recently, liposome preparation steps have simplified by using microfluidic devices (${\mu}FDs$) where liposomes are formed when a stream of lipids in solvent is hydrodynamically focused between two oblique buffer streams in a microchannel. Herein, we report a simple method for the production of immunoliposomes (rabbit IgG-conjugated liposomes) using microvalve-controlled ${\mu}FD$. The presence of antibody on the liposome was verified by observing the binding of immunoliposomes to rabbit IgG on the surface. The results suggest that immunoliposomes can be easily prepared through sequential mixing of antibody, conjugation reagents, preformed liposomes using microvalve-controlled ${\mu}FD$.

• Journal of Microbiology and Biotechnology
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• v.17 no.10
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• pp.1629-1637
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• 2007

An immunochromatography (ICG) strip test using a nanocolloidal gold-antibody probe was developed and optimized for the rapid detection of aflatoxin B1 (AFB1). A monoclonal antibody specific to AFB1 was produced from the cloned hybridoma cell (AF78), coupled with nanocolloidal gold, and distributed on the conjugate pad of the ICG strip test. The visual detection limit of the ICG strip test was 0.5 ng/ml, and this method showed a cross-reaction to aflatoxin B2, G1, and G2. In total, 172 grain and feed samples were collected and analyzed by both the ICG strip test and HPLC. The results of the ICG strip test showed a good agreement with those obtained by HPLC. These results indicated that the ICG strip test has a potential use as a rapid and cost-effective screening tool for the determination of AFB1 in real samples and could be applied to the preliminary screening of mycotoxin in food and agricultural products, generating results within 15 min without complicated steps.

• Korean Journal of Microbiology
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• v.32 no.4
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• pp.280-284
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• 1994

Lack of lysosomal fusion with symbiosomes in symbiont-bearing Amoeba proteus may be due either to the presence of a component in the symbiosome membrane or to the absence of a component needed in the fusion process. Using monoclonal antibody as a probe, lipopolysaccharides were identified as symbiosome-membrane components contributed by symbionts and were found to be exposed on the cytoplasmic side of the membrane. In order to test whether lipopolysaccharides may play a role in the prevention of lysosome-symbiosome fusion, the antilipopolysaccharides antibody was microinjected and processed for double immunostaining in conjuction with anti-lysosome antibody as a lysosome-fusion indicator. Microinjection of the anti-LPS antibody caused symbiosomes to fuse with lysosomes, suggesting that X-bacterial lipopolysaccharides could be 'fusion-preventing' factors.

• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.4
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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.

• The Korean Journal of Zoology
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• v.34 no.1
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• pp.44-53
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• 1991

Distribution of the antigens during the cell cycle of amoebae was followed by immunof-luorescence microscopy using a monoclonal antibody against the nucleus as a probe. While the cells were in the interphase, the antigen was localized on the nucleus membrane. But it was dispersed all over the cytoplasm during mitosis and cytokinesis. The molecular weights of the immunoreacted antigens were 210 KD and 190 KD as determined by SDS PAGE and western blotting of the purified nuclei. The antigens were not soluble in non-ionic detergent, but were released from the nucleus by incubation with 0.05 M sodium carbonate, pH 10.6 or with 8 M urea at serial chemical extraction. Thus the antigens appeared to be peripheral proteins of the nurBeus envelope. The isoelectic point of both antigens was 7.64 as determined by 2 D PAGE and transfer blotting. Considering the peiipherd association with the nucleus membrane and the dispersed distribution during mitosis, the antigens could be lamin like proteins. Hourever, it appears also possible that they are the component molecules of the unusually structured aurous lamina of amoeba nucleus since they have the large molecular weight and the basic pl.

• Animal cells and systems
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• v.3 no.2
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• pp.215-219
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• 1999

Monoclonal antibody for delta-9-tetrahydrocannabiol (THC Ab), conjugated with protein A-gold, was employed as a probe to detect THC localization in the gland and subjacent cells of chemically fixed bracts of Cannabis. THC was detected in the outer wall of the disc cells, fibrillar matrix, the surface feature of secretory vesicles, and sheath throughout development of the secretory cavity. The probe was absent from vesicles. Label was also present in anticlinal walls of disc cells and walls of dermal and mesophyll cells. Little or no THC Ab was present in disc cells and none were detected in control tissues. This distribution pattern of THC Ab was similar to that in tissues prepared by high pressure cryofixation-cryosubstitution. Consistent association of THC with wall and wall-derived materials suggests that cannnabinoids are synthesized outside the plasma membrane and bound to a wall component, where-upon they are transported to the cavity with wall materials released from the disc cell wall during development of the secretory cavity.

• Journal of Electrochemical Science and Technology
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• v.14 no.1
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• pp.51-58
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• 2023

The effective detection of human chorionic gonadotropin (HCG) is considerably important for the clinical diagnosis of both of early pregnancy and nonpregnancy-related diseases. In this work, a simple and sensitive electrochemical sandwich-type immunosensing platform was designed by synthesizing b-cyclodextrin (CD) functionalized graphene (CD/GN) hybrid as simultaneously sensing platform and signal transducer coupled with rhodamine b (RhB) as probe. In brief, GN offers large surface area and high conductivity, while CD exhibits superior host-guest recognition capability, thus the primary antibody (Ab1) of HCG can be bound into the cavities of CD/GN to form stable Ab1/CD/GN inclusion complex; meanwhile, the secondary antibody (Ab2) and RhB can also enter into the cavities, producing RhB/Ab2/CD/GN complex. Then, by using Ab1/CD/GN as sensing platform and RhB/Ab2/CD/GN as signal transducer (in which RhB was signal probe), a simple sandwich-type immunosensor was constructed. Under the optimum parameters, the designed immunosensor exhibited a considerable low analytical detection of 1.0 pg mL^-1 and a wide linearity of 0.002 to 10.0 ng mL^-1 for HCG, revealing the developed sandwich-type electrochemical immunosensing platform offered potential real applications for the determination of HCG.

• IMMUNE NETWORK
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• v.2 no.4
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• pp.227-232
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• 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.