• Journal of Microbiology
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• v.45 no.6
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• pp.547-552
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• 2007

Previously, we generated monoclonal antibodies (MAbs) that bound to the surface of human embryonic stem cells (hESCs) in an attempt to discover new hESC-specific surface markers. In this study, MAb 47-235 (IgG1, ${\kappa}$) was selected for further characterization. The MAb bound to the surface of undifferentiated hESCs but did not bind to mouse ESCs or mouse embryonic fibroblast cells in flow cytometric analysis. The antibody immunoprecipitated a 47 kDa protein from the lysates of cell surface-biotinylated hESCs. Identification of the protein by quadrupole time of flight tandem mass spectrometry revealed that 47-235 binds to Ag 243-5 protein of Mycoplasma arginini. BM-Cyclin treatment of the hESCs that reacted with 47-235 resulted in loss of mycoplasma DNA and the reactivity to 47-235. Nevertheless, the hESCs that were reactive to 47-235 maintained self-renewal and pluripotency and thus could be differentiated into three embryonic germ layers.

• Journal of Radiopharmaceuticals and Molecular Probes
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• v.6 no.1
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• pp.39-45
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• 2020

Recently, single-domain antibodies (sdAb) are bioengineered for molecular imaging applications. Single-domain antibody, obtained from naturally occurring antibodies in camelid species and cartilaginous fish is the smallest fully functional antigen-binding antibody fragments of heavy-chain. Since their discovery, they have been investigated extensively in clinical therapeutics, monitoring and diagnostics. Their small size is important advantage for high solubility, high stability, fast blood clearance and rapid targeting. This review article summarizes the recent status of this new antibody to visualize, diagnose or inhibit specific targets of cancer.

• Bulletin of the Korean Chemical Society
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• v.27 no.4
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• pp.479-483
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• 2006

We developed a microfluidic immunoassay platform for the detection of various analytes such as bacterial pathogen by packing antibody-immobilized glass beads in spatially-isolated microchambers on a microfluidic device. Primary amines of antibody were covalently conjugated to carboxyl-terminated glass beads previously treated with aminosilane followed by glutaraldehyde. Through this covalent binding, up to 905 $\mu$g immunoglobulin G (IgG) per gram of glass beads was immobilized. For application, glass beads attaching antibody specific to Escherichia coli O157:H7, a foodborne pathogen, were packed into a microfluidic device and used for the detection of the serotype. This prototype immunoassay device can be used for the simultaneous detection of multiple analytes by sequentially packing different-sized glass beads attaching different antibody in discrete microchambers on a single microfluidic device.

• Journal of Microbiology and Biotechnology
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• v.15 no.3
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• pp.579-586
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• 2005

When subjected to hyperosmotic pressure by NaCl addition, H69K-NGD transfectoma, like KR12H-2 transfectoma, displayed decreased specific growth rate (${\mu}$) and increased specific antibody productivity ($q_{Ab}$): Elevation of medium osmolality from 280 mOsm/kg to 415 mOsm/kg decreased ${\mu}$ by $79\%$ in batch cultures of H69K-NGD transfectoma, while it increased $q_{Ab}$ by $103\%$. However, unlike KR12H-2 tranfectoma, enhanced $q_{Ab}$ of H69K-NGD transfectoma at hyperosmolalities was not due to elevated levels of Ig mRNAs. In hyperosmotic cultures of H69K-NGD transfectoma, heavy-chain mRNA per cell was not enhanced with increasing osmolality. Hyperosmotic pressure was found to preferentially enhance immunoglobulin (Ig) translation rates of H69K-NGD transfectoma. However, under hyperosmotic pressure, the translation rate of Ig polypeptides was not enhanced as much as $q_{Ab}$. This result suggests that hyperosmotic pressure also influences the post-translational process. Taken together, the results obtained show that intracellular response of transfectomas to hyperosmotic pressure, in regard to the main intracellular steps of the antibody secretory pathway, is cell-line dependent.

• Archives of Pharmacal Research
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• v.18 no.5
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• pp.308-313
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• 1995

This study has been undertaken to examine the acetylcholinesterase (AChE) of electric organ from korean electtric ray(Narke japonica). Korean electric ray was caughted at Chungmu sea and transported to the laboratory, where electric organs were removed and stored at $-70^{\circ}C$ until used. Acelycholinesterase(AChE) of electric organ was purified by affinity column that was prepared with dicaproyl-methylpyridinium linked to Sepharose 4B. Upon purification, the specific activities in Ellman unit were increased by 52 and 39 times for high salt soluble AChE (HSSE, 870.86 $\DeltaOD/min/geam$ of tissue) and detergent soluble AChE(DSE, 105.42 .$\DeltaOD/min/geam$ of tissue), respectively. Each subunit of AChE separated by SDS polyacrylamide gel electrophoresis(SDS-PAGE)was transferred to immonilon P by western boltting and detected by mAbs raised against each subunit of AChE from electric organ og Torpedo califomica. Collagenic tails of AChE from Torpedo califomica, likewise 103Kd protein of AChE from Narke japonica was detected by monoclonal antibody specific to 103Kd of AChE from Torpedo califomica. However, molar ratio of three subunits of AChE from Narke japonica is different from that of Torpedo calicormica. Furthermore, catalytic subunit of AChE from Narke japonica was not identified by monoclnal antibody specific to catalytic subunit of AChE from Torpedo californica. These results showed differences in molecular structure of AChE from Narke japonica and AChE from Torpedo califormica eventhough they showed same enzymatic activities.

• Clinical and Experimental Pediatrics
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• v.58 no.3
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• pp.89-95
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• 2015

Purpose: House dust mite (HDM) has been suggested to be the most important aeroallergen responsible for atopic asthma in Korea. We aimed to investigate that specific IgE antibodies to HDM and other common indoor aeroallergens contribute differently to total serum IgE and show different relationships with longitudinal fractional exhaled nitric oxide (FeNO) measurements in Korean atopic asthmatic patients. Methods: A total of 193 children aged 8 to 16 years with intermittent or mild persistent atopic asthma were recruited. Sera were assayed for total IgE and specific IgE antibodies to HDM and other common indoor allergens. FeNO was serially measured 10 times or more over 2 years when subjects were not receiving controller medications. Results: In 152 children who completed the study, IgE antibodies to specific HDM were more prevalent than those to other common indoor aeroallergens. In addition, IgE antibody titers to HDM were the strongest contributor to total IgE increases. Furthermore, only HDM-specific IgE antibody titer significantly correlated with maximum FeNO (r=0.21, P=0.029) and the rate of FeNO higher than 21 parts per billion (ppb) (r=0.30, P=0.002). Eight patients (5%) were found to have maximum FeNO of 21 ppb or less, suggesting the presence of a low FeNO phenotype among atopic asthmatic patients. Conclusion: The quantity of HDM-specific IgE antibody provides a possible explanation for increases of total IgE and significantly correlates with the amount and frequency of FeNO increases in Korean atopic asthmatic patients.

• Journal of Animal Science and Technology
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• v.56 no.1
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• pp.3.1-3.7
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• 2014

We examined the effects of Prunella vulgaris Labiatae (P. vulgaris L.) on specific and non-specific immune responses of Nile tilapia, Oreochromis niloticus. The optimal concentration without toxicity of P. vulgaris was determined to $30-40{\mu}g/ml$ in vitro and $120{\mu}g$/100 g of fish in vivo. P. vulgaris significantly elicited an antibody titer compared to FCA or ${\beta}$-glucan. ${\beta}$-glucan plus P. vulgaris group synergistically enhanced antibody production. No significant difference in antibody production was observed between P. vulgaris and P. vulgaris plus ${\beta}$-glucan group. A respiratory burst activity of head kidney (HK) leucocytes of tilapia administered with 300 or $500{\mu}g$ P. vulgaris was significantly (p < 0.05) enhanced compared with the PBS-injected control group and FCA-treated group. Maximum increase in the NBT reduction value was observed in $500{\mu}g$ P. vulgaris group but no significant difference was found between 300 and $500{\mu}g$ P. vulgaris group. The level of serum lysozyme activity was significantly (p < 0.05) higher in the 300 and $500{\mu}g$ P. vulgaris than $100{\mu}g$ P. vulgaris and FCA group. The phagocytic activities of HK leucocytes from tilapia administered with 300 and $500{\mu}g$ P. vulgaris were significantly (p < 0.05) higher than $100{\mu}g$ P. vulgaris and the control group. P. vulgaris was revealed with a good immunoadjuvant evoking the specific and non-specific immune responses of tilapia.

• Journal of fish pathology
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• v.23 no.1
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• pp.9-16
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• 2010

We determined the effects of temperature shifts on the kinetics of the numbers of antibody-secreting cell (ASC) in the olive flounder Paralichthys olivaceus immunised with formalin-killed Edwardsiella tarda. When fish that were acclimated to $22^{\circ}C$ and immunised at that temperature were transferred to a lower temperature ($12^{\circ}C$) at a various times (immediately, 1, 2 or 4 weeks) after immunisation, both further differentiation of B cells and secretion of antibody from the ASC developed at $22^{\circ}C$ were suppressed at $12^{\circ}C$. However, in the converse experiment ($12^{\circ}C$ to $22^{\circ}C$), the magnitude of the humoral immune response was recovered independent of the time of the transfer after immunisation at low temperature, even though the peak levels of each transferred group did not reach the level found in $22^{\circ}C$ control group. The results were confirmed by counting the number of specific antibody secreting cells (SASC) in the spleen. This study provides the evidences of the immune reaction that the potential for antibody production in B cells of flounder, the most important species in aquatic industry of Korea, immunized at high temperature is suppressed by subsequent exposure to low temperature and that low temperature-induced humoral immuno suppression can be reversed by exposure to a higher temperature.

• Toxicological Research
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• v.27 no.2
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• pp.125-131
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• 2011

Through the present study, we produced a monoclonal antibody against aflatoxin B1 (AFB1) using AFB1-carboxymethoxylamine BSA conjugates. One clone showing high binding ability was selected and it was applied to develop a direct competitive ELISA system. The epitope densities of AFB1-CMO against BSA and KLH were about 1 : 6 and 1 : 545, respectively. The monoclonal antibody (mAb) from cloned hybridoma cell was the IgG1 subclass with ${\lambda}$-type light chains. The $IC_{50}s$ of the monoclonal antibody developed for AFB1, AFB2, AFG1 and AFG2 were 4.36, 7.22, 6.61 and 29.41 ng/ml, respectively, based on the AFB1-KLH coated ELISA system and 15.28, 26.62, 32.75 and 56.67 ng/ml, respectively, based on the mAb coated ELISA. Cross-relativities of mAb to AFB1 for AFB2, AFG1 and AFG2 were 60.47, 65.97 and 14.83% in the AFB1-KLH coated ELISA, and 59.41, 46.66 and 26.97% in the mAb coated ELISA, respectively. Quantitative calculations for AFB1 from the AFB1-Ab ELISA and AFB1-Ag ELISA ranged from 0.25 to 25 ng/ml ($R^2$ > 0.99) and from 1 to 100 ng/ml ($R^2$ > 0.99), respectively. The intra- and inter-assay precision CVs were < 10% in both ELISA assay, representing good reproducibility of developed assay. Recoveries ranged from 79.18 to 91.27%, CVs ranged from 3.21 to 7.97% after spiking AFB1 at concentrations ranging from 5 to 50 ng/ml and following by extraction with 70% methanol solution in the Ab-coated ELISA. In conclusion, we produced a group specific mAb against aflatoxins and developed two direct competitive ELISAs for the detection of AFB1 in feeds based on a monoclonal antibody developed.

• Biomolecules & Therapeutics
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• v.2 no.4
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• pp.303-309
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• 1994

The analysis of membrane receptors for hormones and neurotransmitters has progressed considerably by pharmacological and biochemical means and more recently through the use of specific antibodies. Two kinds of antibodies could be produced, one is from synthetic peptides and the other from proteins such as purified receptor. Anti-peptide antibodies gave some advantages; epitope is evident and also receptor purification in quantity is not prerequisite. It can be also applied to the study of receptor structure-activity relationship. The purpose of the present study was 1) to produce and characterize a polyclonal antibody against a synthetic $\beta$2-adrenergic receptor peptide(Phe-Gly-Asn-Phe-Trp-Cys-Phe-Trp-Thr-Ser-Ile-Asp-Val-Leu) and 2) to determine the effects of this antibody on the $\beta$-adrenergic receptor ligand interaction. The peptide sequence contains an amino acid residue such as Asp-113 which was identified as one of important component for receptor-ligand interaction in site-directed mutagenesis studies. Production of antibody was performed by immunization of rabbits through popliteal lymph node with the peptide coupled with Keyhole Limpet Hemocyanin (KLH). The titer of antibody against this peptide was 1 : 1000. The anti-peptide antibody was able to detect a 67 kDa protein band in western blot corresponding to the molecular weight of the $\beta$-adrenergic receptor in partially purified receptor fraction derived from guinea pig lung. The antisera inhibited the specific binding of [$^3$H]dihydroalprenolol to $\beta$-adrenergic receptor in a concentration-dependent manner. The results from this study suggest that the peptide sequence selected in the present study is important for the receptor ligand interaction.