• 한국작물학회지
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• 제34권s01호
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• pp.40-47
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• 1989

In spite of the development of highly sophisticated instrument, the precise quantitation of plant hormones still has many difficulties. Due to their high specificity, sensitivity and minimal sample purification steps, immunological assays have been widely applied for plant hormone assay. Enzme-linked immunosorbent assay technique for the determination of plant hormones was developed by Voller in 1978. Immunological assays are accomplished by competition of labeled tracer antigen and unlabeled antigen for a limited number of specific antibodies. The use of enzyme as replacement labels for radioisotopes enabled much of the sensitivity and specificity of radioimmunoassay (RIA) to be retained but without the inherent disadvantage of high capital cost, potential health hazard, and short shelf life of the labeled reactants.

• 약학회지
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• 제31권2호
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• pp.70-81
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• 1987

To elucidate action mechanism of lyophyllan A, an antitumor polysaccharide of Lyophyllum decastes, its immunological activities were examined. Lyophyllan A increased significantly the weights of spleen and liver of mice. Lyophyllan A also restored the decreased thymic weight in tumor-bearing mice. It did not show any direct cytotoxicity against tumor cells, but showed immunopotentiating activities by increasing the number of the plaques in hemolytic plaque assays. Lyophyllan A increased the number of peritoneal exudate cells (PEC) and inhibited the growth of sarcoma 180 mixed with PEC. Moreover the macrophages from lyophyllan A-treated mice exhibited a strong cytotoxic activity towards L5178Y target cells.

• IMMUNE NETWORK
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• 제21권6호
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• pp.39.1-39.18
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• 2021

The high virulent severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus that emerged in China at the end of 2019 has generated novel coronavirus disease, coronavirus disease 2019 (COVID-19), causing a pandemic worldwide. Every country has made great efforts to struggle against SARS-CoV-2 infection, including massive vaccination, immunological patients' surveillance, and the utilization of convalescence plasma for COVID-19 therapy. These efforts are associated with the attempts to increase the titers of SARS-CoV-2 neutralizing Abs (nAbs) generated either after infection or vaccination that represent the body's immune status. As there is no standard therapy for COVID-19 yet, virus eradication will mainly depend on these nAbs contents in the body. Therefore, serological nAbs neutralization assays become a requirement for researchers and clinicians to measure nAbs titers. Different platforms have been developed to evaluate nAbs titers utilizing various epitopes sources, including neutralization assays based on the live virus, pseudovirus, and neutralization assays utilizing recombinant SARS-CoV-2 S glycoprotein receptor binding site, receptor-binding domain. As a standard neutralization assay, the plaque reduction neutralization test (PRNT) requires isolation and propagation of live pathogenic SARS-CoV-2 virus conducted in a BSL-3 containment. Hence, other surrogate neutralization assays relevant to the PRNT play important alternatives that offer better safety besides facilitating high throughput analyses. This review discusses the current neutralization assay platforms used to evaluate nAbs, their techniques, advantages, and limitations.

• 한국어업기술학회:학술대회논문집
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• 한국어업기술학회 2000년도 춘계수산관련학회 공동학술대회발표요지집
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• pp.446-446
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• 2000

Specific polyclonal and/or monoclonal antibodies (MAbs) to immunoglobulins (Igs) and their subunits have proved to be valuable tools in immunological research and in immunological assays. In this study, we developed and characterized MAbs against flounder serum Igs. To obtain the pure flounder serum Igs, mouse IgG (mIgG) was immunized to flounder. Flounder Igs were purified by using mIgG-agarose affinity column chromatography. The structure of purified flounder Ig was observed, on denatured SDS-PAGE, to be composed of two heavy chains (77 and 72 kd) and two light chains (28 and 26 kd). MAbs were produced by fusion of myeloma cells (SP2/0) with Balb/c mouse spleen cells previously primed with the flounder Igs. Finally, three hybridoma clones, FIM 511, FIM 519 and FIM 562 were established to recognize both 2 heavy chains, 26 kd of light chain and 28 kd of light chain, respectively. On the other hand, the flounder immune sera collected on the weekly basis were tested on ELISA and immunoblot analysis whether boosting effect is present in flounder humoral immune system. As a result, the secondary immune response in flounder was ascertained on ELISA, but not on immunoblot analysis. Further, we observed an alteration of serum protein levels following immunization. Our MAbs and basic information on flounder humoral immune system obtained in this study will be helpful to control and monitor the efficiency of fish vaccines and therapeutic process of flounder diseases.

• 한국작물학회지
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• 제48권5호
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• pp.407-412
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• 2003

In order to measure an antifreezing tolerance, antifreeze proteins accumulated upon cold acclimation in apoplast were analyzed. As Dongborilho were cold-acclimated for 3 to 74 days there was an abrupt increase in apoplastic proteins up to 30 days and then decrease to the similar levels. Among the known antifreeze proteins, CLP produced in E. coli. and TLP purified from apoplast were used to generate antisera that allow to measure and localize the proteins in leaves of barley. The CLP of 27.7 kDa and TLPs of 6, 26, 27, 30, and 31 kDa were increased in their amounts in apoplast as cold treatment being longer. There was a correlation among the amounts of those proteins accumulated in apoplast and freezing tolerance as shown in field and ion leakage tests for five cultivars. The deposit of CLP was localized in the marginal area and the area adjacent to leaf vescular bundle cells in an increasing manner according to duration of cold acclimation but no variation was observed in terms of it's distribution. Based on the close correlation between levels of antifreeze proteins and degrees of freezing tolerance, the immunological methods was to develop to estimate a freezing tolerance in barley.

• Journal of Veterinary Science
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• 제21권4호
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• pp.59.1-59.12
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• 2020

Background: Atopic dermatitis (AD) is a common chronic inflammatory skin disease. To understand AD, there have been many trials establishing AD animal models. Although various trials to establish AD animal models have been existed, even the mechanisms of AD in animal models are not enough clarified. Objectives: This study assessed AD characteristics induced in Nishiki-nezumi Cinnamon/Nagoya (Nc/Nga) mice following trinitrochlorobenzene (TNCB) treatment for different periods and house dust mite (HDM) treatment to compare each model's immunological patterns, especially with cytokine antibody array tool. Methods: In this study, we exposed Nc/Nga mice to TNCB or HDM extract to induce AD. Nc/Nga mice were divided into 4 groups: control, TNCB 2 weeks-treated, TNCB 8 weeks-treated, and HDM-treated groups. After AD induction, all mice were evaluated by serum immunoglobulin E (IgE) concentration and serum cytokine antibody assays, scoring of skin lesions, scoring of scratching frequency, and histological analysis. Results: The results showed significant differences between groups in serum IgE concentration, skin lesion scores, and scratching frequency. The analysis results for serum cytokine antibody arrays showed that in the TNCB 8 weeks- and HDM-treated groups, but not in the TNCB 2 weeks-treated group, expressions of genes related to the immune response were enriched. Among the histological results, the skin lesions in the HDM-treated group were most similar to those of AD. Conclusions: We confirmed that immunological pattern of AD mice was markedly different between HDM and TNCB treated groups. In addition, the immunological pattern was quietly different dependent on TNCB treated duration.

• Interdisciplinary Bio Central
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• 제2권2호
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• pp.1.1-1.8
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• 2010

Microdevices have been used as effective experimental tools for the rapid and multiplexed analysis of individual cells in single-cell assays. Technological advances for miniaturizing such systems and the optimization of delicate controls in micron-sized space homing cells have motivated many researchers from diverse fields (e.g., cancer research, stem cell research, therapeutic agent development, etc.) to employ microtools in their scientific research. Microtools allow high-throughput, multiplexed analysis of single cells, and they are not limited by the lack of large samples. These characteristics may significantly benefit the study of immune cells, where the number of cells available for testing is usually limited. In this review, I present an overview of several microtools that are currently available for single-cell analyses in two popular formats: microarrays and microfluidic microdevices. Then, I discuss the potential to study human immunology on the single-cell level, and I highlight several recent examples of immunoassays performed with single-cell microdevice assays. Finally, I discuss the outlook for the development of optimized assay platforms to study human immune cells. The development and application of microdevices for studies on single immune cells presents novel opportunities for the qualitative and quantitative characterization of immune cells and may lead to a comprehensive understanding of fundamental aspects of human immunology.

• Food Science and Biotechnology
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• 제18권1호
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• pp.89-94
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• 2009

Frequent outbreaks of foodborne illness have been increasing the awareness of food safety. Conventional methods for pathogen detection and identification are labor-intensive and take days to complete. Some immunological, rapid assays are developed, but these assays still require prolonged enrichment steps. Recently developed biosensors have shown potential for the rapid detection of foodborne pathogens. In this study, an impedimetric biosensor was developed for rapid detection of Salmonella entritidis in food sample. To develop the biosensor, an interdigitated microelectrode (IME) was fabricated by using a semiconductor fabrication process. Anti-Salmonella antibodies were immobilized based on neutravidin-biotin binding on the surface of the IME to form an active sensing layer. To evaluate the effect of electrode gap on sensitivity of the sensor, 3 types of sensors with different electrode gap sizes (2, 5, and $10{\mu}m$) were fabricated and tested. The impedimetric biosensor could detect $10^3\;CFU/mL$ of Salmonella in pork meat extract with an incubation time of 5 min. This method may provide a simple, rapid, and sensitive method to detect foodborne pathogens.

• 한국어병학회지
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• 제28권2호
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• pp.93-98
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• 2015

Full-length recombinant VP2 and VP3 proteins of aquabirnavirus isolated from olive flounder were expressed successfully in E. coli expression system. After rats were immunized with these proteins, antisera were used for in vitro and in vivo neutralization test. In in vitro test, VP2 antibody titers were higher than that of VP3. In in vivo assays, fish challenged with aquabirnavirus neutralized with VP2 antibody survived longer than other fish.

• Journal of Microbiology and Biotechnology
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• 제21권5호
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• pp.529-535
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• 2011

Single-chain variable fragment (scFv) is a fusion protein of the variable regions of the heavy (VH) and light (VL) chains of immunoglobulin, connected with a short linker peptide of 10 to about 20 amino acids. In this study, the scFv of a monoclonal antibody against the third domain of human CD4 was cloned from OKT4 hybridoma cells using the phage display technique and produced in E. coli. The expression, production, and purification of anti-CD4 scFv were tested using SDS-PAGE and Western blot, and the specificity of anti-CD4 scFv was examined using ELISA. A 31 kDa recombinant anti-CD4 scFv was expressed and produced in bacteria, which was confirmed by SDS-PAGE and Western blot assays. Sequence analysis proved the ScFv structure of the construct. It was able to bind to CD4 in quality ELISA assay. The canonical structure of anti-CD4 scFv antibody was obtained using the SWISS_MODEL bioinformatics tool for comparing with the scFv general structure. To the best of our knowledge, this is the first report for generating scFv against human CD4 antigen. Engineered anti-CD4 scFv could be used in immunological studies, including fluorochrome conjugation, bispecific antibody production, bifunctional protein synthesis, and other genetic engineering manipulations. Since the binding site of our product is domain 3 (D3) of the CD4 molecule and different from the CD4 immunological main domain, including D1 and D2, further studies are needed to evaluate the anti-CD4 scFv potential for diagnostic and therapeutic applications.