• The Journal of the Korean Society for Microbiology
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• v.20 no.1
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• pp.91-102
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• 1985

The advantages of enzyme-linked immunosorbent assay(ELISA) are its senstivity and simplicity in detecting IgG, IgM and IgA antibody. To apply ELISA to diagnosis of typhoid fever, antigen such as lipopolysaccharide of Salmonella typhi or killed whole cell must be coated on solid phase. It is easy to coat lipopolysaccharide on ELISA plate but troublesome to purify it. As it is easy to obtain the killed whole cells, the development of the appropriate method by which those antigens of S. typhi are optimally coated on solid phase is needed. To establish the appropriate method, carbonate buffer, methanol or poly-L-lysine was applied as binding substance on polystyrene or polyvinylchloride plate as solid phase when the killed whole cell antigens of S. typhi varided as follows: $10^6$, $10^7$, $10^8$ and $10^9\;cell/ml$. The criteria of the optimal method were determined as follows: 1. The optical density of positive sera is above 1.0(0.6 in IgM) at 1:10 serum dilution and is 0.3(0.2 in IgM) higher than that of negative sera: 2. The O.D. of sera is flat or lowering according to serum dilution: 3. It must be that the O.D. of negative sera is lower than 0.2 at the point of serum dilution where the O.D. of positive sera is higher than 1.0(0.5 in IgM). The results obtained were summarized as follows: 1. The methods which fitted the above criteria were to use poly-L-lysine as binding substance, polyvinylchloride plate as solid phase and $10^7\;cell/ml$ as antigen concentration of S. typhi(poly-L-lysine/polyvinylchloride/$10^7$) and poly-L-lysine/polyvinylchloride/$10^8$ in detecting IgG antibody, methanol/polystyrene/$10^9$, poly-L-lysine/polyvinylchloride/$10^8$ and poly-L-lysine/polyvinylchloride/$10^9$ in IgM and carbonate buffer/polystyrene/$10^8$, carbonate buffer/polystyrene/$10^9$, methanol/polystyrene/$10^8$, methanol/polyvinylchloride/$10^8$, methanol/polyvinylchloride/$10^9$, poly-L-lysine/polyvinylchloride/$10^8$ and poly-L-lysine/polyvinylchloride/$10^9$ in IgA. 2. The coaling method using poly-L-lysine, polyvinylchloride plate and $10^8\;cell/ml$ was best to assay IgG, IgM and IgA antibody all in one. By this method, to assay the each immunoglobulin calss with an appropriate fixed serum dilution, 1:320 dilution was best.

• The Korean Journal of Physiology
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• v.23 no.1
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• pp.89-98
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• 1989

In order to produce antibody for use in radioimmunoassay of gastrin in physiological concentration, four rabbits of New Zealand white were immunized with synthetic human gastrin-17-I conjugated to hemocyanin with EDC. Among them, only one rabbit produced antibody that could bind 50% of $^{125}I-gastrin$ at a final dilution of 1:25,000. $^{125}I-gastrin$ was prepared with synthetic human gastrin-17-I and $NaI^{125}$ by lactoperoxidase technique. The product was then purified on a column of Sephadex Gl5/G5O (7:3, w/w) followed by a column of DEAE sephadex A-25. The specific radioactivity of the purified $^{125}I-gastrin$ was in the range of 347-1429 ${\mu}Ci/nmole$ when determined by the self-displacement method. The effective affinity constant $(K_{eff})$, total binding sites (N), heterogeneity index $({\alpha})$ and average affinity constant $(K_{0})$ of the anti-gastrin serum calculated from Scatchard plot as well as Sips plot were $1.77{\times}10^{11}/M$, 255 nM, 0.84 and $0.79{\times}10^{11}/M$, respectively. When radioimmunoassay was performed with the anti-gastrin serum, it was confirmed that the mean concentration of gastrin immunoreactivity in plasma was increased by feeding in humans and rats, and also increased by bombesin administration in rats. The results indicate that the anti-gastrin serum produced in the present investigation is suitable for radioimmunological determination of gastrin in physiological concentration.

• Parasites, Hosts and Diseases
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• v.25 no.1
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• pp.1-6
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• 1987

This study was to observe the changes of blastogenic responses of splenic Iymphocytes to T-cell mitogens, N. fcwleri Iysate and concanaualin A, and serum antibody titer during the course of experimental PAM in mice. Naegleria fcwleri, strain 0359, was cultured in the CGVS medium axenically and inoculated intranasally with $7{\times}10^4$ trophozoites for the development of experimental PAM in mice. The amoebae were subjected to ultrasonication and centrifuged at 20,000g for 60 minutes, and filtered through $0.2{\mu\textrm{m}}$ filter membrane. The supernatant, N. fcwleri Iysate, was used as T-cell mitogen, and antigen for ELISA. The serum antibody was examined by ELISA using peroxidase conjugate. Two hundred ${\mi}l$ of $10^6$ splenocytes in RPMI 1640 containing 0% fetal calf serum were added to each well of a microtiter plate. To each well was added T-cell mitogens, $100{\mu}g/ml$ of N. fowleri Iysate or $4{\mu}g/ml$ of con. A, and the plates were incubated for 42 hours at $37^{\circ}C$ in 5% $CO_2$ incubator. Cultures were pulsed with of $methyl-(^3H)-thymidine$ 6 hour before harvesting. The mean blastogenic response of the splenocytes to N. fewleri Iysate was reduced, whereas that to con. A was also reduced up to on day 11 after infection. Both of these results were statistically significant compared with those of uninfected control group. The serum antibody titers were increased gradually up to day 15. The results indicated that there was an impairment of the blastogenic response of splenocytes to N. fowleri Iysate during the acute course of experimental PAM in mice.

• Applied Chemistry for Engineering
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• v.30 no.4
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• pp.429-434
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• 2019

A lateral flow immunoassay platform utilizing antibody functionalized water soluble CdSe/ZnS semiconductor quantum dots (QDs) was developed for the analysis of human serum amyloid A-1 (hSAA1) in a buffer solution. hSAA1 was chosen as a target protein because it is regarded as a potential biomarker associated with early diagnosis and prognosis in patients of lung cancer. The immunoassay strip on a nitrocellulose membrane was fabricated by spraying two lines composed of a test line with a monoclonal antibody against hSAA1 (10G1) (anti hSAA1) and a control line of anti-chicken IgY. While the CdSe/ZnS QDs synthesized in an organic phase were transferred to a water phase by ligand exchange using carboxylic acid modified alkane thiol. The QDs was then conjugated to monoclonal antibody against hSAA1 (14F8) [anti hSAA1 (14F8)] and used as a fluorescent detection probe. The sequential lateral flow of hSAA1 in different concentration and QDs-anti hSAA1 (14F8) complex allowed to form the surface sandwich complex of anti hSAA1 (10G1)/hSAA1/QD-anti hSAA1 (14F8), which was then analyzed using fluorescence microscope. A 100 nM concentration of hSAA1 protein can be detected by naked eyes under an optimized lateral flow buffer condition with a sensing time of 5 mins.

• Journal of radiological science and technology
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• v.10 no.1
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• pp.85-90
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• 1987

Preliminary studies on the measurement of the free thyroxine in the serum with Amerlex $FT_{4}$ RIA kit were investigated using a tracer as $^{125}I-T_{4}$ derivative which is not almostly bound to thyroxine binding globulin, etc. The results are followed: 1. Linearity was tested on standards at various concentrations, and reproducibility and accuracy was excellent. 2. The antibody specificity is also excellent, and standard calibration curve of total $T_{4}$ was similar that of adding the TBG inhibitor. 3. Each value of $T_{4}$ in serum (the normal group, the hypothyroidism patients the pregnant women and the TBG dificiency patients) was not significant. As mentioned above, this method is more simple and rapid, compared to the other method.

• Journal of Life Science
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• v.9 no.1
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• pp.35-39
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• 1999

Elecrophoreticl analysis of a 36 kDa protein was runned by SDS-PAGE, isoelectric focusing (IEF) and two dimensional electrophoresis pattern. Major 36 kDa and 25, 46, 48, 66 kDa protein were detected by Coomassie blue stain on SDS-PAGE. Major 36kDa protein was eluted for production of antiserum for serological analysis, IEF and two dimensional electrophoresis. Isoelectric point of 36kDa was aout pH 8.5. Two dimensional electrophoresis of eluted 36kDa showed one point on the gel. Anti-36 kDa serum made by newzilland rabbit for serological test. In ELISA, final titer of antibody was 100×{TEX}$2^5}${/TEX} : 1. Neutralize ability of serum was examined by slide agglutination test and colonization test in rat. Anti-36 kDa serum agglutinated whole cell of V. vulnificus were inhibited colonization on intestine in rat. Accordingly In this paper contain some electrophoretical analysis and serological test of a 36 kDa OMP of V. vulnificus.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.21 no.1
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• pp.38-54
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• 2008

Objective : This study was designed to investigated effects of Pinelliae Rizoma (PR) on asthma Methods : Detecting antigen specific antibody isotypes, cytokines in serum and bronchoalveolar lavage fluid (BALF). In addition, the present author also investigated changes in weight of spleen and proliferation rates of splenocytes. Finally, histopathological observation of the lung tissue was also investigated. Results : Oral administration of PR lowered OVA specific IgE levels in serum, IgG1 levels in serum and BALF. PR also decreased production levels of IL-4 in BALF. In addition, total cells in BALF were decreased by oral administration of PR effectively. In histapathological observation, PR group showed downward tendency of inflammatory cell infiltration around small vessels. Conclusion : PR is useful to treat patients with asthma and the mechanisms are related in suppression of Th2 skewing reactions.

• Microbiology and Biotechnology Letters
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• v.17 no.5
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• pp.481-488
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• 1989

A serum-free medium that could be used for the large-scale culture of mouse hybridoma to produce monoclonal antibodies was developed. The medium was based on a 1:1 mixture of Iscove's Modified Dulbecco's Medium and Ham's F-12, supplemented with insulin 10$\mu\textrm{g}$/$m\ell$, transferrin 10$\mu\textrm{g}$/$m\ell$, ethanolamine 10$\mu$M and selenium 30nM (designated EBM (enriched basal medium) with the supplements). The effect of various supplements of steroid hormones, vitamins, lipid and mineral salts was investigated and their optimal concentration was determined to replace fetal calf serum (PCS). These components were added respectively and then added by way of two or three combination to discern of which component combination was effective to the culture of hybridoma. As a result, serum-free medium KM3 (EBM with BSA 100$\mu\textrm{g}$/$m\ell$, mineral cocktail and 0.05% PEG) was deter-mined. The hybridoma Alps 25-3 cultured in this medium showed almost the same growth rate as in medium added with 2% fetal bovine serum. However, the antibody concentration from KM3 cultures was 80% of that obtained from culture with FCS. KM3 was also examined for the culture of other mouse hybridomas, KW, A4W & HCGK, and it was confirmed that it could support the growth of these hybridomas and the production of monoclonal antibodies.

• Korean Journal of Veterinary Service
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• v.43 no.3
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• pp.173-179
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• 2020

The objective of this study was to evaluate the usefulness of detection of PRRSV and PRRSV-specific antibodies in oral fluids for monitoring of PRRSV infection in endemic farms. The level of PRRSV and anti-PRRSV antibodies in serum and oral fluids was evaluated in five age groups of pigs (6, 9, 12, 16 weeks of age and gilts). The samples (25 serums and 5 oral fluids/per a farm) were collected from 5 different farms endemically infected by PRRSV. Both serum and oral fluid samples were tested for PRRSV by quantitative reverse-transcription polymerase chain reaction (qRT-PCR) and for anti-PRRSV antibodies by two commercial PRRSV ELISA kits. ELISA mean s/p ratios (2.98 vs 1.63) and positive rate (84.0% vs 68.8%) of the oral fluid samples showed significantly higher levels but had similar patterns to the seroprofile of the blood samples. The PRRSV positive rate of oral fluid and serum samples was 40.0% and 44.0% respectively. In conclusion, the use of oral fluids for PRRS monitoring in endemic farms is strongly recommended.

• Microbiology and Biotechnology Letters
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• v.18 no.1
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• pp.61-65
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• 1990

Lectin related inducer can enhance IgG$_1$ production rate from murine hybridoma cells by employing step-feeding of serum free media with producing about 40 mg/$\ell$ of monoclonal antibodies. This step-feeding perfusion process also proves to be able to cutivate animal cells when serum free media can not support the growth of these cells in perfusion process, as well as to improve production rate. This process yields about 28 x 10$^{-10}$ mg of MAb/cells/h compared to 11.1 x 10$^{-10}$ and 4.0 x 10$^{-11}$ mg/cells/h for perfusion process and batch cultivation with 10% serum containing media, respectively.