• Title/Summary/Keyword: protein antigenicity

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Reduction of Allergic Potential of Meju by Three Step Fermentation (3단계 발효에 의한 콩 알레르기성의 저하)

  • Ryu, Chung-Ho;Lee, Jeong-Ok;Son, Dae-Yeul
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.8
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    • pp.1066-1071
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    • 2012
  • In this study we investigated the change in antigenicity and allergenicity of Meju, a traditional Korean soybean product, by fermentation via 3 different microorganisms. The steamed soybeans were fermented with Lactococcus lactis subsp. lactis and/or Aspergillus oryzae and/or Bacillus subtilis. Proteins in soybean were degraded after fermentation. Antigenicity or allergenicity were analysed by immunoblotting and ELISA using soybean protein-specific polyclonal antibodies or soybean allergic patient sera. The best degradation was achieved by three step fermentation using nisin-producing Lactococcus lactis subsp. lactis IFO12007, A. oryzae and B. subtilis. Allergenicity and antigenicity were also starkly reduced after three step fermentation. The three-step fermentation method developed in our lab suggests an excellent alternative to reduce the allergenicity of soybeans.

Comparison of antigenicity of Edwardsiella tarda isolates in loach(Misgurnus mizloepis) (미꾸라지에서의 Edwardsiella tarda isolates의 항원성 비교)

  • Lee, Young;Jun, Lyu-Jin;Kim, Myoung-Suk;Park, Kyung-Hyun;Jeong, Hyun-Do
    • Journal of fish pathology
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    • v.21 no.3
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    • pp.201-208
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    • 2008
  • We compared the pathogenicity and antigenicity of two different Edwardsiella tarda (E. tarda) strains KFE and Edk-2 isolated in Korea and Japan respectively. In the pathogenicity analysis with challenge test against loach, E. tarda KFE isolate showed stronger pathogenicity compared to that of E. tarda Edk-2 isolate. The differences were also confirmed by the comparison of OMP (outer membrane protein) in SDSPAGE which showed three major bands, 41kDa, 37kDa and 30kDa, for E. tarda KFE isolates and two major bands, 41kDa and 30kDa, for E. tarda Edk-2 isolates. On the base of these results, we tried to determine the differences of antigenicities of these two isolates in loach which is one of the important species in freshwater aquaculture in Korea. Numbers of specific antibody secreting cells (SASC), appeared to be higher in loach immunized with FKC of E. tarda Edk-2 than loach immunized with FKC of E. tarda KFE. ELISPOT-assay for the comparison of antigenicity showed relatively high percentage of cross-reaction and implied the presence of some common epitopes in the antigens of these two E. tarda isolates.

Phylogenetic analysis and antigenic determinant prediction of red sea bream iridovirus isolated in Korea from 2019 to 2023 (2019년부터 2023년까지 국내에서 분리된 참돔이리도바이러스의 계통 분류 및 항원 결정기 예측)

  • Guk Hyun Kim;Joon Gyu Min;Hyun Do Jeong;Kwang Il Kim
    • Journal of fish pathology
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    • v.37 no.1
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    • pp.25-36
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    • 2024
  • In this study, we analyzed the phylogenetic classification, epitope prediction, and pathogenicity of red sea bream iridovirus (RSIV) isolated from rock bream between 2019 and 2023. Phylogenetics based on genes encoding MCP and ATPase indicated that all five RSIV isolates belonged to RSIV subtype II. The deduced amino acid sequence of the MCP for the amplicons (1362 bp) obtained from RSIV isolates had a length of 453 amino acids. Among these, the amino acid sequences of the RSIV-19, 21, 22, and 23 isolates showed 100% identity, while the RSIV-20 isolate showed 99.78% identity with one residue difference at position 306. As a result of antigenicity analysis based on amino acid sequence, the antigenicity score of the RSIV-20 isolate was 0.6386 and the other RSIV isolates were 0.6365. Additionally, the prediction of their antigenic determinants resulted in a total of 17 identical antigenic plots. When each RSIV was inoculated into rock bream, no significant differences were observed with 100% cumulative mortality in all groups. This study provides data on the potential for genetic variation of RSIV isolated in the same marine area over the past five years, and the antigenicity and pathogenicity results of each isolate are expected to be useful information for selecting future vaccine strains.

Detection of Human Anti-Trypanosoma cruzi Antibody with Recombinant Fragmented Ribosomal P Protein

  • Kim, Yeong Hoon;Yang, Zhaoshou;Lee, Jihoo;Ahn, Hye-Jin;Chong, Chom-Kyu;Maricondi, Wagner;Dias, Ronaldo F.;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.57 no.4
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    • pp.435-437
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    • 2019
  • Chagas disease is caused by the protozoan parasite Trypanosoma cruzi, and is endemic in many Latin American countries. Diagnosis is based on serologic testing and the WHO recommends two or more serological tests for confirmation. Acidic ribosomal P protein of T. cruzi showed strong reactivity against positive sera of patients, and we cloned the protein after fragmenting it to enhance its antigenicity and solubility. Twelve positive sera of Chagas disease patients were reacted with the fragmented ribosomal P protein using western blot. Detection rate and density for each fragment were determined. Fragments F1R1, F1R2, and F2R1 showed 100% rate of detection, and average density scoring of 2.00, 1.67, and 2.42 from a maximum of 3.0, respectively. Therefore, the F2R1 fragment of the ribosomal P protein of T. cruzi could be a promising antigen to use in the diagnosis of Chagas disease in endemic regions with high specificity and sensitivity.

Experimental study of antigenicity test of Sweet Bee Venom in Guinea Pigs (기니픽을 이용한 Sweet Bee Venom의 항원성 평가)

  • Cho, Byung-Jun;Kwon, Ki-Rok
    • Journal of Pharmacopuncture
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    • v.14 no.4
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    • pp.23-32
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    • 2011
  • Objectives: This study was performed to examine the antigenic potential of pure melittin (Sweet Bee Venom - SBV) extracted from the bee venom by utilizing protein isolation method of gel filtration. Methods: All experiments were conducted at Biotoxtech (Chungwon, Korea), authorized a non-clinical studies institution, under the regulations of Good Laboratory Practice (GLP). Antigenic potential of SBV was examined by active systemic anaphylaxis (ASA) and passive cutaneous anaphylaxis (PCA) in guinea pigs. SBV was subcutaneously administered at 0.07 and 0.28mg/kg and also as a suspension with adjuvant (Freund's complete adjuvant: FCA). Ovalbumin (OVA) as a suspension with adjuvant was used to induce positive control response ($5mg/m{\ell}$-FCA). Results: 1. In the ASA test, experimental groups showed some symptoms of anaphylaxis like piloerection, hyperpnea and staggering gait. 2. In the PCA test, low dosage group did not show any antibody responses, whereas high dosage group showed positive responses. 3. In the weight measurement and clinical observation, experimental groups didn't show any significant changes compared with control group. 4. In the autopsy of body, the abnormalities of lung were detected in the corpse. This means that the cause of death may induced anaphylactic shock. Conclusions: Above findings suggested that SBV had antigenic potential in guinea pig. Further studies on the subject should be conducted to yield more concrete evidences.

Antigenicity of Recombinant Human G-CSF (CJ-50001) (CJ-50001(rG-CSF)에 대한 항원성시험)

  • Baek, Nam-Jin;Kang, Jae-Ku;Kim, Dal-Hyun;Mok, K.-Hun;Kim, Je-Hak;Kim, Hyun-Su
    • Toxicological Research
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    • v.13 no.3
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    • pp.303-306
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    • 1997
  • Antigenic potential of genetically engineered human granulocyte colony-stimulating factor (CJ-50001) was assessed in guinea pigs and mice. In active systemic anaphylaxis (ASA) test, although CJ-50001 at 50 $\mu\textrm{g}$ /head induced anaphylactic responses, CJ-50001 5 $\mu\textrm{g}$ /head alone or 50 $\mu\textrm{g}$ / head with adjuvant did not induce anaphylactic responses. In passive systemic anaphylaxis test (PCA) or passive hemagglutination test (PHA), CJ-50001 did not induce positive responses. It is concluded that, in light of the fact that CJ-50001 was antigenic only in ASA but not in PCA or PHA and also that CJ-50001 is a foreign human recombinant protein to guinea pigs, CJ-50001 may not induce systemic allergic react-ion in its clinical use in human.

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Antigenicity of Partial Fragments of Recombinant Pasteurella multocida Toxin

  • Lee, Jeong-Min;Woo, Hee-Jong
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1756-1763
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    • 2010
  • Pasteurella multocida serogroup D strain, which produces P. multocida toxin (PMT), is a widespread and harmful pathogen of respiratory diseases such as pneumonia and progressive atrophic rhinitis (PAR) in swine. Vaccination has been considered the most desirable and effective approach for controlling the diseases caused by toxigenic P. multocida. To investigate the antigenicity and immunogenicity of partial fragments of recombinant PMT, recombinant proteins of the N-terminal (PMT-A), middle (PMT-B), C-terminal (PMT-C), and middle-C-terminal (PMT2.3) regions of PMT were successfully produced in an Escherichia coli expression system. The molecular masses of PMT-A, PMT-B, PMT-C, and PMT2.3 were ca. 53, 55, 35, and 84 kDa, respectively, purified by nickel-nitrilotriacetic acid (Ni-NTA) affinity column chromatography. All the recombinant proteins except for PMT-A showed immune responses to antisera obtained from a swine showing symptoms of PAR. Moreover, high titers of PMT-specific antibodies were raised from mice immunized with each of the recombinant proteins; however, the immunoreactivities of the antibodies to authentic PMT and heat-inactivated whole bacteria were different, respectively. In the protection study, the highest protection against homologous challenge was shown in the case of PMT2.3; relatively poor protections occurred for the other PMT fragments.

Molecular cloning of ribosomal P protein in Toxoplasma gondii and the availability to detect antibody against recombinant protein in toxoplasmosis patients

  • Ahn, Hye-Jin;Kim, Sera;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.41 no.2
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    • pp.89-96
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    • 2003
  • Among the panel of monoclonal antibodies (mAb) against Toxoplasma gondii, mAb of Tg621 (Tg621) clone blotted 38 kDa protein which localized in the cytoplasm of tachyzoites by immunofluorescence microscopy The protein was not released into the parasitophorous vacuole during or after invasion. The cDNA fragment encoding the protein was obtained by screening a T. gondii cDNA expression library with Tg621. The full length cDNA sequence was completed with 5’-RACE as 1,592 bp, which contained open reading frame of 942 bp. The deduced amino acid sequence of Tg621 consisted of a polypeptide of 313 amino acids, with significant homology to ribosomal P proteins (RPP) of other organisms especially high to those of apicomplexan species. The expressed and purified TgRPP was assayed in western blot with the sera of toxoplasmosis patients and normal sera, which resulted in the 74.0% of positive reactions in toxoplasmosis patients whereas 8.3% in normal group. Therefore, the antibody formation against TgRPP in toxoplasmosis patients was regarded as specific for T. gondii infection and suggested a potential autoantibody.

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.

Allergenicity of Soybeans Depending on Their Variety (품종에 따른 콩의 알레르기성)

  • Son, Dae-Yeul;Kim, Ye-Jin
    • Korean Journal of Food Science and Technology
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    • v.42 no.5
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    • pp.627-631
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    • 2010
  • The allergenicity of soybeans was analyzed using polyclonal antibodies and the blood sera of patients with soybean allergy, using fourteen different varieties of soybeans that are consumed in Korea. The study that used polyclonal antibodies having specificity for soybeans showed that while some of the fourteen varieties of soybeans contained additional protein bands indicating antigenicity, others lacked such bands, and the most antigenic protein was found in Jinpum soybeans. In comparing blood sera reactivity of four patients having soybean allergies, who had antigenicity values of 65U/ml or more according to CAP testing, the soybean varieties of Danbaek and Shinpaldal2 had the most reactivity and Daewon had the least. The result that the allergenicity of proteins in soybeans differs according to the variety of soybean, leads to the conclusion that it may be possible to reduce consumer allergic reactions to soybean products by choosing an appropriate variety of soybeans.