• Title/Summary/Keyword: Antibody Engineering

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Pharmacological, Toxicological Studies of Antitumor Polysaccharides Obtained from Ganoderrna lucidurn IY 009 (Ganoderma lucidum IY 009로 부터 분리된 항암성 다당류의 약리 및 독성)

  • Lee, Kweon-Haeng;Lee, Chong-Ock;Lee, June-Woo;Jeong, Hoon;Han, Man-Deuk;Jeong, June-Ho;Oh, Doo-Hwan
    • Microbiology and Biotechnology Letters
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    • v.22 no.2
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    • pp.182-189
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    • 1994
  • The highest antitumor activity was observed in water soluble AS fraction of the Ganoderma lucidum IY 009. AS fraction did not show any cytotoxicity on sarcoma 180 cell but stimulated antibody production, opsonization of macrophage in ICR mouse and superoxide ion production from isolated macrophage. AS fraction activated complement C3 in human serum, and their antitumor activity was inhibited by EDTA, a chelator of cation related complementary activation. AS fraction exerted om prolong of life span and ingibition of tumor growth in the leukemia P388 or L1210 transplanted inbreed mouse,k BDF1 but krestin did not. AS fraction did not show any serious and lethal effects through oral administration on ICR mouse, and LD$_{50}$ of those was above 2,230 mg/kg.

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Acute Phase Protein Lipocalin-2 Is Associated with Formalin-induced Nociception and Pathological Pain

  • Jha, Mithilesh Kumar;Jeon, Sangmin;Jin, Myungwon;Lee, Won-Ha;Suk, Kyoungho
    • IMMUNE NETWORK
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    • v.13 no.6
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    • pp.289-294
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    • 2013
  • Lipocalin-2 (LCN2) is an acute-phase protein induced by injury, infection, or other inflammatory stimuli. LCN2 binds small hydrophobic ligands and interacts with cell surface receptor to regulate diverse cellular processes. The role of LCN2 as a chemokine inducer in the central nervous system (CNS) has been previously reported. Based on the previous participation of LCN2 in neuroinflammation, we investigated the role of LCN2 in formalin-induced nociception and pathological pain. Formalin-induced nociceptive behaviors (licking/biting) and spinal microglial activation were significantly reduced in the second or late phase of the formalin test in Lcn2 knockout mice. Likewise, antibody-mediated neutralization of spinal LCN2 attenuated the mechanical hypersensitivity induced by peripheral nerve injury in mice. Taken together, our results suggest that LCN2 can be therapeutically targeted, presumably for both prevention and reversal of acute inflammatory pain as well as pathological pain.

Uptake of Fibroin Microspheres by 3T3 Cells (3T3 세포에 의한 피브로인 마이크로스피어의 흡수)

  • Lee, Jin Sil;Go, Nam Kyung;Lee, Shin Young;Hur, Won
    • KSBB Journal
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    • v.29 no.5
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    • pp.328-335
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    • 2014
  • Vehicle toxicity is one of the main obstacles for intracellular delivery of bioactive compounds. Silk fibroin is a natural polymer proven to have high biocompatibility since being used as suture material. In this report, fibroin microspheres were prepared without any chemical modification or cross-linking not to affect its biocompatibility. The microspheres were taken up by more than 90% of 3T3 cells. Cellular uptake continued after medium replenishment with a different-colored fluorescent microsphere, suggesting that simultaneous ingestion and exocytosis occurred. Cellular uptake of fibroin microspheres did not affect cell viability. Intracellular trafficking of the microspheres using lysosome-specific fluorescent dye revealed that fibroin microspheres were localized both in the cytoplasm and in the lysosome. Accordingly, fibroin microspheres can be a potential vehicle for intracytoplasmic delivery of large cargos, such as mixtures of proteins, nutrients or artificial organelles.

Preparation and Characteristics of Phosphatidylcholine Liposomes Covalently Coupled with Immunoglobulin Fragments (이뮤노글로부린 토막과 결합된 포스파티딜콜린 리포솜의 제조와 특성)

  • Park, Jung-Hwan;Lee, Eun-Ok;Kim, Jong-Duk
    • Journal of Pharmaceutical Investigation
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    • v.22 no.2
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    • pp.115-124
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    • 1992
  • The characteristics and stabilities of phosphatidylcholine liposomes covalently coupled with immunoglobulin fragments prepared by the REV method were investigated by the dynamic light scattering, absorbance and calcein release. Using a sulfhydryl-reactive phospholipid derivative of N-[4$({\rho}-maleimido-phenyl)$ butyl] phosphatidylethanolamine (MPB-PE), Fab' antibody fragments were covalently combined with preformed large unilamellar vesicles (LUV), Coupling ratio was $250\;{\mu}g$ of $Fab'/{\mu}mol$ of phospholipid in vesicles, From dynamic light scattering, it was found that the size of the vesicles increases as the ratio of cholesterol to lipid increases, but that apparently, the size of liposomes was not sensitive to the existence of Fab' fragments. Regardless of inserting Fab' fragments, the absorbance of liposomes decreased as the amounts of bile salt (BS) added. At very low BS concentrations, BS/lipid aggregates would be formed in the outer vesicles monolayer, while, at the high BS concentrations, mixed micelles would be preferred. The vesicles incorporated with Fab' fragments, however, are more resistant to the bile salts than the MPB-PE vesicle are. The absorbance of vacant liposomes and calcein release resulted in that the Fab' vesicles and MPB-PE vesicles by the REV method are very stable, but that those by the sonication method sufferred the significant change of turbidities.

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Fiber-Optic Sensor Simultaneously Detecting Localized Surface Plasmon Resonance and Surface-Enhanced Raman Scattering

  • Norov, Erdene;Jeong, Hyeon-Ho;Park, Jae-Hyoung;Lee, Seung-Ki;Jeong, Dae Hong
    • Rapid Communication in Photoscience
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    • v.2 no.2
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    • pp.46-51
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    • 2013
  • This study reports a fiber-optic sensor detecting biomolecule by simultaneously monitoring localized surface plasmon resonance (LSPR) from gold nanoparticles (Au NPs) of ca. $50{\pm}5$ nm attached on one end of optical fiber and surface enhanced Raman scattering (SERS) of the reporter molecules adsorbed on the gold surfaces as an additional sensing tool. The sensor was fabricated by immobilizing Au NPs on one end of an optical fiber by chemical reaction. LSPR and SERS signals of the sensor were measured using various refractive indices solutions. Finally, the sensor was applied to observe real-time LSPR sensor-gram and SERS spectra of the reporter molecule of 4-aminothiphenol during the antibody-antigen reaction of interferon-gamma (IFN-${\gamma}$) as a proof-concept experiment of biological applications.

Characterization and Identification of Bacillus thuringiensis subsp. tenebrionis SR6 and SR8

  • Kim, Il-Gi;Lee, Jae-Wook;Suh, Suk-Chul;Rhim, Seong-Lyul
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.772-776
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    • 2004
  • Physiological and molecular characteristics of Bacillus thuringiensis SR6 and SR8 were investigated, and phase contrast and electron microscopies revealed that a large rhomboidal crystal protein was present in the sporulating cells. SDS-PAGE and Western blot analyses showed that B. thuringiensis SR8 produced 70 kDa protein much more than other proteins, and that the 70 kDa protein could bind to the antibody of B. thuringiensis subsp. tenebrionis-crystal toxin protein, indicating that the crystal 70 kDa protein has an immunological homology with B. thuringiensis subsp. tenebrionis-crystal toxin protein. The DNA fragment of B. thuringiensis subsp. tenebrionis-toxin gene was detected in B. thuringiensis SR6 and SR8 by using PCR amplification analysis. Furthermore, the insect bioassay showed the insecticidal activity against Colorado potato beetle larvae. Based on the physiological and molecular similarities to B. thuringiensis subsp. tenebrionis, it is suggested that the B. thuringiensis SR6 and SR8 may be mutants of the B. thuringiensis subsp. tenebrionis strain overexpressing the crystal of 70 kDa toxin protein.

Glyco-engineering of Biotherapeutic Proteins in Plants

  • Ko, Kisung;Ahn, Mi-Hyun;Song, Mira;Choo, Young-Kug;Kim, Hyun Soon;Ko, Kinarm;Joung, Hyouk
    • Molecules and Cells
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    • v.25 no.4
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    • pp.494-503
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    • 2008
  • Many therapeutic glycoproteins have been successfully generated in plants. Plants have advantages regarding practical and economic concerns, and safety of protein production over other existing systems. However, plants are not ideal expression systems for the production of biopharmaceutical proteins, due to the fact that they are incapable of the authentic human N-glycosylation process. The majority of therapeutic proteins are glycoproteins which harbor N-glycans, which are often essential for their stability, folding, and biological activity. Thus, several glyco-engineering strategies have emerged for the tailor-making of N-glycosylation in plants, including glycoprotein subcellular targeting, the inhibition of plant specific glycosyltranferases, or the addition of human specific glycosyltransferases. This article focuses on plant N-glycosylation structure, glycosylation variation in plant cell, plant expression system of glycoproteins, and impact of glycosylation on immunological function. Furthermore, plant glyco-engineering techniques currently being developed to overcome the limitations of plant expression systems in the production of therapeutic glycoproteins will be discussed in this review.

Radiation dosimetry of 89Zr labeled antibody estimated using the MIRD method and MCNP code

  • Saeideh Izadi Yazdi ;Mahdi Sadeghi ;Elham Saeedzadeh ;Mostafa Jalilifar
    • Nuclear Engineering and Technology
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    • v.55 no.4
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    • pp.1265-1268
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    • 2023
  • One important issue in using radiopharmaceuticals as therapeutic and imaging agents is predicting different organ absorbed dose following their injection. The present study aims at extrapolating dosimetry estimates to a female phantom from the animal data of 89Zr radionuclide accumulation using the Sparks-Idogan relationship. The absorbed dose of 89Zr radionuclide in different organs of the human body was calculated based on its distribution data in mice using both MIRD method and the MCNP simulation code. In this study, breasts, liver, heart wall, stomach, kidneys, lungs and spleen were considered as source and target organs. The highest and the lowest absorbed doses were respectively delivered to the liver (4.00E-02 and 3.43E-02 mGy/MBq) and the stomach (1.83E-03 and 1.66E-03 mGy/MBq). Moreover, there was a good agreement between the results obtained from both MIRD and MCNP methods. Therefore, according to the dosimetry results, [89Zr] DFO-CR011-PET/CT seems to be a suitable for diagnostic imaging of the breast anomalies for CDX-011 targeting gpNMB in patients with TNBC in the future.

Identification of a conservative site in the African swine fever virus p54 protein and its preliminary application in a serological assay

  • Xu, Lingyu;Cao, Chenfu;Yang, Zhiyi;Jia, Weixin
    • Journal of Veterinary Science
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    • v.23 no.4
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    • pp.55.1-55.12
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    • 2022
  • Background: ASF was first reported in Kenya in 1910 in 1921. In China, ASF spread to 31 provinces including Henan and Jiangsu within six months after it was first reported on August 3, 2018. The epidemic almost affected the whole China, causing direct economic losses of tens of billions of yuan. Cause great loss to our pig industry. As ELISA is cheap and easy to operate, OIE regards it as the preferred serological method for ASF detection. P54 protein has good antigenicity and is an ideal antigen for detection. Objective: To identify a conservative site in the African swine fever virus (ASFV) p54 protein and perform a Cloth-enzyme-linked immunosorbent assay (ELISA) for detecting the ASFV antibody in order to reduce risks posed by using the live virus in diagnostic assays. Method: We used bioinformatics methods to predict the antigen epitope of the ASFV p54 protein in combination with the antigenic index and artificially synthesized the predicted antigen epitope peptides. Using ASFV-positive serum and specific monoclonal antibodies (mAbs), we performed indirect ELISA and blocking ELISA to verify the immunological properties of the predicted epitope polypeptide. Results: The results of our prediction revealed that the possible antigen epitope regions were A23-29, A36-45, A72-94, A114-120, A124-130, and A137-150. The indirect ELISA showed that the peptides A23-29, A36-45, A72-94, A114-120, and A137-150 have good antigenicity. Moreover, the A36-45 polypeptide can react specifically with the mAb secreted by hybridoma cells, and its binding site contains a minimum number of essential amino acids in the sequence 37DIQFINPY44. Conclusions: Our study confirmed a conservative antigenic site in the ASFV p54 protein and its amino acid sequence. A competitive ELISA method for detecting ASFV antibodies was established based on recombinant p54 and matching mAb. Moreover, testing the protein sequence alignment verified that the method can theoretically detect antibodies produced by pigs affected by nearly all ASFVs worldwide.

Expression and Localization of Brain Glutamate Dehydrogenase with Its Monoclonal Antibody

  • Lee, Jong-Eun;Choe, Su-Yeong;Jo, Seong-U
    • Animal cells and systems
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    • v.2 no.1
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    • pp.71-80
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    • 1998
  • Glutamate dehydrogenase (GDH) is one of the main enzymes involved in the formation and metabolism of the neurotransmitter glutamate. In the present study, we investigated the distribution of the GDH-immunoreactive cells in the rat brain using monoclonal antibodies against bovine brain GDH isoprotein. GDH-immunoreactive cell were distributed in the basal ganglia, thalamus and the nuclei belong to substantia innominata, and its connecting area, subthalamic nucleus, zona incerta, and substantia niqra. We could see GDH-immunoreactive cells in the hippocampus, septal nuclei associated with the limbic system, the anterior thalamic nuclei connecting between the hypothalamus and limbic system, and its associated structures, amygdaloid nuclear complex, the dorsal raphe and median raphe nuclei and the reticular formation of the midbrain. The GDH-immunoreactive cells were shown in the pyramidal neurons of the cerebral cortex, the Purkinie cells of the cerebella cortex, their associated structures, ventral thalamic nuclei and the reticular thalamic nuclei that seem to function as neural conduction in the thalamus.

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