• Journal of Periodontal and Implant Science
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• v.38 no.4
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• pp.565-578
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• 2008

Purpose: Porphyromonas gingivalis(P. gingivalis) heat shock protein (HSP)60 may play a role in the immunopathogenesis of periodontitis as well as atherosclerosis by modulating autoimmune reaction due to its high level of sequence homology between bacteria and human counterpart. The purpose of this study was to identify immunodomiant epitope of P. gingivalis HSP60 that is reactive exclusively to the homologous bacteria without reacting with human HSP. Materials and methods: The present study was performed to identify the peptide specifically recognized by anti-P. gingivalis HSP60 monoclonal antibodies mono-reactive to P. gingivalis HSP60. Results: Four different hybridomas were cloned producing monoclonal IgG antibodies exclusively to P. gingivalis HSP60. Thirty seven synthetic peptides (20-mer with 5-amino acid overlapping) were synthesized. All of these peptide were subject to SDS-PAGE for immunblot analysis. One peptide (TVPGGGTTYIRAIAALEGLK) and the other peptide (TLVVNRLRGSLKICAVKAPG) were recognized by all and one of the four monoclonal antibodies, respectively, that reacted solely with P. gingivalis HSP60. Immunohistochemistry to identify the localization of the HSP60 in the diseased gingival tissues revealed that all of the four monoclonal antibodies were highly reacted with the diseased gingival tissue than normal gingival tissue. Conclusion: The P. gingivalis HSP60 peptides (TVPGGGTTYIRAIAALEGLK and TLVVNRLRGSLKICAVKAPG, respectively) are positively involved in the immunopathologic process of periodontal disease. The peptide may potentially be developed as vaccine candidates. Further investigations are under way to identify more clones producing monoclonal antibodies reactive to P. gingivalis HSP and to other periodontopathogenic bacteria as well, while maintaining specificities to human counterpart.

• BMB Reports
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• v.28 no.1
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• pp.27-32
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• 1995

Polyclonal antibodies against human DNA-dependent RNA polymerase II (HPP II) were generated from chicken egg yolk after immunization with RNA polymerase II as an antigen. The antibodies from egg yolk (IgY) were purified and characterized. IgY showed a specificity against DNA-dependent RNA polymerase II, and was a polyclonal antibody against 12 subunits of polymerase II. An amount of 0.35 mg of IgY was obtained freman HPP II-Sepharose affinity column using 10 eggs from a chicken immunized against RNA polymerase II as an antigen. These antibodies can be used for isolating the genes for RNA polymerase II components, and for in vitro transcription assays using HP-RNA polymerase II.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.829-832
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• 2001

Phage display technique as a new antibody production method can express the protein on the minor coat of phage particle in a library constructed by utilizing a recombination of genes coding the variable regions of immunoglobulin. This new method is particularly advantageous in producing antibodies against toxic substances and compounds with low immunogenicities. We first confirmed the concept of antibody expression on the phage particle by selecting a positive control of the phage library (e.g., Griffin.l donated from MRC center in England). The library was then employed to produce antibodies specific to human serum albumin via repetitive bio-panning procedure. The mean affinity of the antibodies selected gradually increased along with the number of bio-panning, which demonstrated that the phage display method could produce monocloanl antibodies with high affinities.

• Parasites, Hosts and Diseases
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• v.51 no.4
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• pp.467-470
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• 2013

The present study reports a human case of cutaneous gnathostomiasis with recurrent migratory nodule and persistent eosinophilia in China. A 52-year-old woman from Henan Province, central China, presented with recurrent migratory reddish swelling and subcutaneous nodule in the left upper arm and on the back for 3 months. Blood examination showed eosinophila (21.2%), and anti-sparganum antibodies were positive. Skin biopsy of the lesion and histopathological examinations revealed dermal infiltrates of eosinophils but did not show any parasites. Thus, the patient was first diagnosed as sparganosis; however, new migratory swellings occurred after treatment with praziquantel for 3 days. On further inquiring, she recalled having eaten undercooked eels and specific antibodies to the larvae of Gnathostoma spinigerum were detected. The patient was definitely diagnosed as cutaneous gnathostomiasis caused by Gnathostoma sp. and treated with albendazole (1,000 mg/day) for 15 days, and the subsequent papule and blister developed after the treatment. After 1 month, laboratory findings indicated a reduced eosinophil count (3.3%). At her final follow-up 18 months later, the patient had no further symptoms and anti-Gnathostoma antibodies became negative. Conclusively, the present study is the first report on a human case of cutaneous gnathostomiasis in Henan Province, China, based on the past history (eating undercooked eels), clinical manifestations (migratory subcutaneous nodule and persistent eosinophilia), and a serological finding (positive for specific anti-Gnathostoma antibodies).

• Food Science and Biotechnology
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• v.14 no.3
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• pp.410-412
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• 2005

Vibrio parahaemolyticus is a gram-negative bacterium which acts as a causative agent for food poisoning. Studies with respect to specific extracellular proteins of V. parahaemolyticus would be useful for the development of specific detection methods against V. parahaemolyticus. In our present study, outer membrane proteins (OMPs) of V. parahaemolyticus were obtained from insoluble traction of 1% sarkosyl treated-cell wall materials. SDS-PAGE analysis showed the presence of several conserved outer membrane proteins among five strains of V. parahaemolyticus, and three bands were identified as V. parahaemolyticus OMPs through MALDI-TOF analysis. Polyclonal antibodies enriched with anti-OmpU were obtained from immunized rabbits. The antibodies against these proteins may be useful for the development of detection methods for V. parahaemolyticus.

• Journal of Nutrition and Health
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• v.29 no.9
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• pp.971-980
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• 1996

The soluble transferrin receptor(TfR) in human serum has been shown recently to be a truncated form of intact membrane bound receptor containing most of the extracellular domain. We purfied the transferin-free TfR from human serum by immounoaffinity chromatography which produced the single protein identity in high resolution gel chormatography. The monoclonal antibodies(MAb) against purifed serum TfR were produced by fusion of spleen cells o fimmunized Balb/c mice and SP2 cells. Ten hybrids producing MAb specific for serum TfR were identifed and determine their iostypes. A immunoraddiometric assay (IRMA) for serum TfR was established using two monoclonal IgG1 antibodies as the coating and indicator antibodies on the bosis of their suitability in sandwich IRMA of serum TfR. The mean serum TfR levels in the 15 normal male, 15 normal female, and 19 iron-deficient subjects were 5.4$\pm$0.98, 4.6$\pm$0/76, and 18.0$\pm$12.8mg/1, respectively, and the difference in mean values between normal and iron deficient subjects was significant(p=0.0005). There existed the inverse logarithmic relationship(r=-0.9336, p<0.0001) between the serum TfR and ferritin levels.

• Clinical and Experimental Pediatrics
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• v.50 no.2
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• pp.143-150
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• 2007

Purpose : This study was conducted to validate enzyme immunoassay (EIA) for the quantitative measurement of human IgG antibodies specific for Haemophilus influenzae type b (Hib) capsular polysaccharide. Method : We evaluated specificity, repeatability, intermediate precision, accuracy, lower limit of quantification (LLOQ), and stability to validate standardized EIA for the quantitative measurement of human anti-polyribosylribitol phosphate (PRP) IgG antibodies. Results : The results indicated that this EIA showed specificity to HbO-HA antigen and repeatability and intermediate precision were within acceptance criteria (repeatability: $CV{\leq}15%$, intermediate precision: $CV{\leq}20%$). The EIA-derived results from this laboratory were equivalent to those obtained by the standard radioactive antigen binding assay (RABA) for quantitation of anti-PRP antibodies in the 28 sera. Spiking recovery result was within acceptance criteria ($100{\pm}20%$). The precision and accuracy of samples in LLOQ were from -14.7 to -4.7% in nominal values, which were within acceptance criteria (precision: $CV{\leq}25%$, accuracy: ${\pm}25%$). Freeze-thaw stability and short term temperature stability were within ${\pm}20%$ of acceptance criteria. Conclusions : The EIA which is performed at the Center for Vaccine Evaluation and Study Ewha Medical Research Institute, is an appropriate serologic assay which can be used for quantitation of anti-PRP IgG antibodies in human sera.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.9
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• pp.4255-4261
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• 2012

The present study was conducted to evaluate radioimmunoimaging (RII) and in vivo distribution of mixed antibodies $^{99m}Tc$-EGFR-mAb and $^{99m}Tc$-CD44-mAb in nude mice bearing human lung adenocarcinoma xenografts. Single and mixed applications of the two radiolabeled monoclonal antibodies (mAbs) were compared. Direct labeling of $^{99m}Tc$ was applied to radiolabel the EGFR and CD44 mAbs. The properties of the radiolabeled antibodies were then characterized. RII and assessment of the distribution of the antibodies in nude mice bearing lung adenocarcinoma xenografts were achieved by applying separate and combined doses of $^{99m}Tc$-EGFR-mAb and $^{99m}Tc$-CD44-mAb. The labeling rates of $^{99m}Tc$ for EGFR-mAb and CD44-mAb were $91.5%{\pm}3.8%$ and $92.3%{\pm}4.1%$ respectively, with specific activities of 2.8 and $2.9MBq/{\mu}g$, respectively, and radiochemical purities (RCP) of 96.5% and 96.2%. The radioactivity uptake of the combined application of both radiolabeled antibodies was clearly higher than with a single application of either alone. The relative values of target-to-nontarget (T/NT) measured through the regional interest (ROI) technique were $5.59{\pm}0.42$ (mixed antibodies), $2.78{\pm}0.20$ ($^{99m}Tc$-EGFR-mAb), and $2.28{\pm}0.16$ ($^{99m}Tc$-CD44-mAb) in the RII. The body distribution of the radiolabeled antibodies and their imaging results were basically identical. Application of the mixed antibodies with $^{99m}Tc$-EGFR-mAb and $^{99m}Tc$-CD44-mAb can increase the radioactivity uptake of tumor tissue, leading to more ideal target-to-nontarget ratios, and therefore superior results.

• BMB Reports
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• v.38 no.6
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• pp.703-708
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• 2005

We cloned and expressed human pyridoxal-5'-phosphate (PLP) phosphatase, the coenzymatically active form of vitamin $B_6$, in Escherichia coli using pET15b vector. Monoclonal antibodies (mAb) were generated against purified human brain PLP phosphatase in mice, and four antibodies recognizing different epitopes were obtained, one of which inhibited PLP phosphatase. The binding affinities of these four mAbs to PLP phosphatase, as determined using biosensor technology, showed that they had similar binding affinities. Using the anti-PLP phosphatase antibodies as probes, we investigated their cross-reactivities in various mammalian and human tissues and cell lines. The immunoreactive bands obtained on Western blots had molecular masses of ca. 33 kDa. Similarly fractionated extracts of several mammalian cell lines all produced a single band of molecular mass 33 kDa. We believe that these PLP phosphatase mAbs could be used as valuable immunodiagnostic reagents for the detection, identification, and characterization of various neurological diseases related to vitamin $B_6$ abnormalities.

• BMB Reports
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• v.39 no.2
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• pp.208-215
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• 2006

The human ribosomal protein S3 (rpS3) was expressed in E. coli using the pET-I5b vector and the monoclonal antibodies (mAbs) were produced and characterized. A total of five hybridoma cell lines were established and the antibodies recognized a single band of molecular weight of 33 kDa on immunoblot with purified rpS3. When the purified rpS3 was incubated with the mAbs, the UV endonuclease activity of rpS3 was inhibited up to a maximum of 49%. The binding affinity of mAbs to rpS3 determined by using a biosensor technology showed that they have similar binding affinities. Using the anti-rpS3 antibodies as probes, we investigated the cross-reactivities of various other mammalian brain tissues and cell lines, including human. The immunoreactive bands on Western blots appeared to be the same molecular mass of 33 kDa in all animal species tested. They also appear to be extensively cross-reactive among different organs in rat. These results demonstrated that only one type of immunologically similar rpS3 protein is present in all of the mammalian brain tissues including human. Furthermore, these antibodies were successfully applied in immunohistochemistry in order to detect rpS3 in the gerbil brain tissues. Among the various regions in the brain tissues, the rpS3 positive neurons were predominantly observed in the ependymal cells, hippocampus and substantia nigra pars compacta. The different distributions of rpS3 in brain tissues reply that rpS3 protein may play an important second function in the neuronal cells.