• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.6
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• pp.552-555
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• 2005

The influence of whole yeast cells $(0{\sim}15%\;w/v)$ on the protein adsorption performance in dye-ligand chromatography was explored. The adsorption of a model protein, bovine serum albumin (BSA), was selected to demonstrate this approach. The UpFront adsorbent $(p=1.5\;g/cm^3)$ derivatised with Cibacron Blue 3GA and a commercially available expanded bed column (20 mm i.d.) from UpFront Chromatography, Denmark, were employed in the batch binding and expanded bed operation. The BSA binding capacity was demonstrated to not be adversely affected by the presence of yeast cells. The dynamic binding capacity of BSA at a $C/C_0=0.1$ biomass concentration of 5, 10, 15% w/v were 9, 8, and 7.5mg/mL of settled adsorbent, respectively.

• The Korean Journal of Nuclear Medicine
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• v.25 no.2
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• pp.259-265
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• 1991

The purpose of this study is to investigate the ability of I-131 labeled polyclonal human immunoglobulin to localize an infection. In our country, indium-111 labeled leukocyte or Tc-99m labeled IgG are not readily available because of compex, time-consuming procedure and cost. So we tried to localize infection with I-131 labeled IgG which could be easily prepared. Six rats, infected with staphylococcus aureus in a thigh muscle, received I-131 labeled IgG intravenously and I-131 labeled bovine serum albumin (BSA) were injected to other 5 infected rats. Scintigrams were made at 1, 4, 24, 48, 72 hour later. The radiopharmaceutical demonstrated significant accumulation at the site of infection. The accumulation of I-131 labeled IgG at the site of infection was significantly (P<0.05) higher than that of I-131 labeled BSA at 48, 72 hour. Similar finding could be found at 24 hour imaging, but it was not significant statistically. Therefore it was found that vascular permeability alone could not account for the mode of action of I-131 labeled IgG and it was considered that specific binding played a role. In conclusion, focal sites of inflammation can be detected with I-131 labeled nonspecific human polyclonal IgG and it seems that this method can also be applied to localization of human infection.

• Korean Journal of Animal Reproduction
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• v.10 no.1
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• pp.109-120
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• 1986

This study was conducted to define the effect of addition of lysolecithin (LC) and 20% v/v rabbit serum to sperm preincubation medium on the induction of acrosome reaction (AR) an fertilizing ability in vitro of LG-added sperm. Ejaculated rabbit sperm from New Zealand White buck was washed once by centrifugation, then preincubated for 2 or 4 hrs in a chemically defined medium (DM), DM plus 20% rabbit serum or BSA-free DM plus 20% rabbit serum at 37$^{\circ}C$ water bath or CO2 incubator. At the end of preincubation LC was added to the preincubated sperm, which was stained at 0.5 to 4 hr later and examined for AR and sperm motility. For in vitro fertilization, gametes were coincubated in DM up to 24 hrs and thereafter fertilized embryos were incubated in BSM -II up to 48 hrs. Addition of LC to 4-hr preincubated sperm was more effective for the AR and sperm motility than that to 2-hr preincubated sperm and optimal concentration of LC for AR was about 80${\mu}$g/ml. A significant increase in AR occured from 20 to 30 min. after addition of 80 to 100${\mu}$g/ml in 4-hr preincubated sperm. BSA-free DM plus 20% rabbit serum showed a higher AR and sperm motility than those of DM plus 20% rabbit serum in LC-added sperm after 4-hr preincubation. The incidence of AR after 4-hr preincubation and at 30 min after 60${\mu}$g/ml LC addition varied greatly among individual bucks. Sixty ${\mu}$g/ml LC-added sperm showed a slight high cleavage rate over control levels, but 100${\mu}$g/ml LC-added sperm showed lower cleavage rate rather than 60${\mu}$g/ml LC. It is concluded that optimal concentration of LC for high AR induction and sperm motility in 4-hr preincubated sperm was about 80${\mu}$g/ml, but 60${\mu}$g/ml level was more useful for in vitro fertilization.

• Analytical Science and Technology
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• v.23 no.6
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• pp.537-543
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• 2010

Dopamine (DA) is an important neurotransmitter molecule of catecholamines. Its deficiency could lead to brain disorder such as Parkinson's disease and schizophrenia. Therefore, it is necessary to establish a suitable analytical technique with sensitivity and simplicity. A competitive enzyme-linked immunosorbent assay for DA has been optimized and characterized. Assay sensitivity is controlled by two factors in competitive immunoassay. One is a nature and concentration of competitor, and the other is those of binder, antibody. Thus, optimization was performed: BSA-DA conjugate and antibody-avidin conjugate were prepared by dual heterobifunctional coupling method using SATA and SMCC. Assay condition was optimized with $6.66\;{\mu}gmL^{-1}$ of BSA-DA and $4.17{\times}10^{-10}\;M$ of antibody-avidin conjugate. A dose-response curve was constructed, and a limit of detection and a dynamic range for DA were accomplished to $2.3{\times}10^{-2}\;{\mu}g\;mL^{-1}$ and four orders of magnitude ($1.0{\times}10^{-7}\;M$ to $1.0{\times}10^{-3}\;M$), respectively. Calibration curve was constructed on dynamic range and least-squares regression of this data gave the following relationship: absorbance = -0.1098 log[DA]+0.0353 ($R^2$ = 0.9956).

• Archives of Pharmacal Research
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• v.20 no.1
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• pp.46-52
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• 1997

To obtain more sensitive immunoassay for methamphetamine (MA) determination, the optimum condition of enzyme-linked immunosorbent assay (ELISA) was investigated in regard to immunogens, antibody purification methods and coating tracers. Activated MA, N-(4-aminobutyl)methamphetamine (4-ABMA), was conjugated with bovine serum albumin (BSA) or keyhole limpet hemocyanin (KLH) and used as immunogen. The antibodies were purified by protein G chromatography or various immunoaffinity chromatography-linked MA-protein ligands, such as MA-BSA, MA-KLH or MA-ovalbumin (OVA). Each purified antibody was characterized by means of sensitivity and cross-reactivity using the three MA-protein coating tracers, MA-BSA, MA-KLH and MA-OVA. The best sensitivity of each antibody was acquired with the MA-OVA tracer although the tracer concentration and the antibody titer level at optimum condition were varied. The antibody with high titer level did not always yield good sensitivity. At optimum condition, immunoaffinity chromatography-purified antibodies were better for sensitivity and for specificity than protein G-purified antibodies. The cross-reactivity of the purified antibodies seemed to be affected by immunogen structure and showed somewhat different patterns according to the immunoaffinity ligand utilized. These data show that the antibody purification method as well as choice of coating tracer and immunogen is essential for the sensitivity and specificity of EIA; the optimum condition for assay should be discovered using various methods and combinations.

• Korean Journal of Veterinary Research
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• v.37 no.3
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• pp.613-618
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• 1997

Direct competitive enzyme linked immunosorbent assay(ELISA) with monoclonal antibodies have been studied for quantitative determination of T-2 toxin from the mold corn. The T-2HS, T-2HS-BSA, T-2HS-HRP and monoclonal antibodies against T-2 toxin produced in the studies were qualified for quantitative ELISA test of T-2 toxin. The mean recovery rate from ground com spiked T-2 toxin was 83%. The meaning range of the T-2 test was 60ng to $2{\mu}g$. According to the recovery results with the com spiked T-2, the tests proved to be suitable in the screening of the moldy feed samples for the presence of T-2 toxin and will be able to become the basis of the ELISA test for the quantitative screening kits of T-2 toxin.

• Journal of Food Hygiene and Safety
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• v.6 no.3
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• pp.111-117
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• 1991

We examined to develop the enzyme linked immunosorbent assay (ELISA) for determination of zearalenone in animal feeds. Zearalenone was first converted to 6'-(carboxymethyl) zearalenone oxime(zearalenone oxime) to get a coupling site and then conjugated to bovine serum albumin(BSA) for use as immunogen and to horseradish peroxidase(HRP) for use as enzyme marker. Antibody against zearalenone was obtained after 11 weeks of immunization of rabbit with zearalenone oxime-BSA. Cross reactivity of the antibody with ${\alpha}-zearalanol,\;{\beta}-zearalenol,\;{\alpha}-zearalanol\;and\;{\beta}-zearalanol$ were 168, 46, 26 and 20% respectiviely. A simple procedure was devised for the screening of zearalenone in feeds using ELISA. Feeds samples(5g) were extracted by blending with 25 ml of methanol-phospate butTered saline-dimethylformate(70 : 29 : 1) and the extract was filtered and aqueous filterate analyzed. It took only 1 hours to do whole procedure for the analysis of zearalenone in feeds by the direct competitive ELISA, and detectable limit was 1-100 ppb. Using this procedure, only 4 of 24 feed samples showed positive results with 3.93-7.43 ppb levels.

• Bulletin of the Korean Chemical Society
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• v.32 no.7
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• pp.2253-2259
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• 2011

The self-assembly of one novel organic complex based on chlorogenic acid (HCA) and 2,2'-bipyridine (2,2'-bipy) has been synthesized and characterized. The complex achieved by hydrogen-bonding interactions, adopted a 1:1 stoichiometry in a solid state. The proton transfer occurred from the carboxyl oxygen to the aromatic nitrogen atom to form salts CA${\cdot}$(2,2'-Hbipy), the 2,2'-Hbipy molecule individually occupies the pseudo-tetragonum that is formed with CA. In this paper, the interactions of CA${\cdot}$(2,2'-Hbipy) with bovine serum albumin (BSA) were studied by fluorescence spectrometry. For CA${\cdot}$(2,2'-Hbipy), HCA and 2,2'-bipy, the average quenching constants for BSA were $2.4384{\times}10^4$, $4.653{\times}10^3$, and $3.059{\times}10^3\;L{\cdot}mol^{-1}$, respectively. The mechanism for protein fluorescence quenching is apparently governed by a static quenching process. The Stern-Volmer quenching constants and corresponding thermodynamic parameters ${\Delta}$H, ${\Delta}$G and ${\Delta}$S were calculated. The binding constants and the number of binding sites were also investigated. The conformational changes of BSA were observed from synchronous fluorescence spectra.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.62-62
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• 2003

This study was carried out to investigate the effects of activation agents on parthenogenetic activation of pig oocytes matured in vitro. The medium used for oocyte maturation was tissue culture medium (TCM) 199 supplemented with 26.19 mM sodium bicarbonate, 0.9 mM sodium pyruvate, 10 $\mu\textrm{g}$/ml insulin, 2 $\mu\textrm{g}$/ml vitamin $B_{l2}$, 25 mM Hepes, 10 $\mu\textrm{g}$/ml bovine apotransferrin, 150 $\mu$M cysteamine, 10 IU/ml PMSG, 10 IU/ml hCG, 10 ng/ml EGF, 0.4% BSA, 75 $\mu\textrm{g}$/ml sodium penicillin G, 50 $\mu\textrm{g}$/ml streptomycin sulfate and 10% pFF. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at 38.5$^{\circ}C$, 5% $CO_2$ in air. Cumulus-free oocytes involving first polar body were activated by exposure to various concentrations of ethanol and exposure time of ethanol in Hepes-buffered NCSU23 medium. Also, oocytes were activated by electric pulse alone or combination with ethanol. For electrical activation, oocytes were rinsed twice in 0.3 M mannitol solution supplemented with 0.1 mM CaC1$_2$, 0.2 mM MgC1$_2$, 0.5 mM Hopes and 0.01% BSA, and transferred to a chamber consisting of two electrodes 1 mm apart which was overlaid with the same activation solution. Oocytes were activated with a single DC pulse of 1.3 ㎸/cm for 30 $\mu$sec. After activation treatments, oocytes were washed three times with Hepes-buffered NCSU23 medium and were washed twice with NCSU23 culture medium containing 0.4% BSA, and then cultured in 500 ${mu}ell$ of the same medium for 20 h at 38.5$^{\circ}C$, 5% $CO_2$ in air. The activation rates of oocytes were higher in 6, 7 and 8% ethanol concentrations compared with 0, 5, 9 and 10% ethanol concentrations. Significantly more oocytes (29.3~33.7%) were activated in the exposure for 8, 10, 12 and 15 min than those in the exposure for 0 and 5 min, but there was no difference due to exposure to 8% ethanol for 8 to 15 min. Electric pulse treatment followed by exposure to ethanol significantly improved the rate of oocyte activation (61.9%) compared with that of other 3 treatments. In conclusion, the optimal activation treatment of ethanol exposure alone for the in-vitro matured pig oocytes was 8% ethanol for 8 to 15 min. Electric pulse treatment followed by ethanol exposure significantly improved the rate of activation.n.

• BMB Reports
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• v.30 no.5
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• pp.352-355
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• 1997

The surface adsorbed protein conformations onto the vaccine adjuvants were observed with a Raman spectroscopy by using the maximum adsorption conditions described previously. The adsorbed state Raman vibrational spectra and subsequent spectral analysis display no conformational changes for BSA or IgG relative to their native species in solution.