• Title/Summary/Keyword: Neutralizing antibody

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Production and characterization of monoclonal antibodies against porcine transmissible gastroenteritis virus (돼지 전염성 위장염 바이러스에 대한 단크론항체 생산 및 특성)

  • Jang, Young-en;Cho, Sun-hee;Kim, Byung-han;Ahn, Jae-moon;Kang, Shien-young
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.336-344
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    • 1998
  • Eight monoclonal antibodies(MAbs) against the transmissible gastroenteritis virus (TGEV) were produced and characterized. Four of the MAbs were produced against a reference TGEV, Purdue strain(P115) and the others were produced against the Korean vaccine virus, Pyungtaek strain. Only one MAb(5C8) produced against P115 had neutralizing activity and was found to be E2 protein-specific. The other seven MAbs(4E2, 4G8, 5H6, 1F8, 2C6, 5H5, and 3A6) had specificity of nucleocapsid protein and no neutralizing activity. All MAbs reacted with different strains of TGEV, but none of the MAbs was reactive with porcine enteropathogenic viruses such as rotavirus, epidemic diarrhea virus and enterovirus by fluorescence antibody(FA) test.

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Production and Characterization of a Recombinant Antibody Neutralizing Botulinum Neurotoxin A (보툴리눔 신경독소 A를 중화하는 재조합 항체의 제조와 특성 분석)

  • Park, Hong-Gyu;Choi, Mieyoung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.1
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    • pp.295-301
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    • 2017
  • Botulinum neurotoxin (BoNT/A) is a neurotoxin that selectively attacks the peripheral cholinergic nerve endings. It is produced by Gram -positive, endospore-forming strict anaerobic bacteria, Clostridium botulinum. Since BoNT/A could be a biothreat agent, as well as a contaminator of food and water supplies, the development of sensitive assays for toxin detection and potent antitoxin for the treatment of intoxication is necessary. In this study, for the purpose of producing monoclonal antibodies (mAbs) that are capable of neutralizing Botulinum neurotoxin type A (BoNT/A), scFv (single-chain variable domain fragment) libraries from the rabbit antisera against BoNT/A was fused to a human IgG. The resulting recombinant scFvIgG antibody protein was expressed in stable cell lines and was purified using a protein A affinity chromatography. The efficacy of scFvIgG mAb was confirmed by ELISA and was evaluated for the neutralization of BoNT/A in vivo. Such an in vivo toxin neutralization assay was performed using mice. Although scFvIgG antibody proteins (10 ug) failed to fully protect the mice challenged with BoNT/A (100,000 $LD_{50}$), it significantly prolonged the survival time. These results suggest that scFvIgG mAb may be capable of neutralizing BoNT/A single-chain variable domain fragment.

Prevalence on protective serum antibodies of canine distemper virus and canine parvovirus in Ulsan area (울산지역의 개 디스템퍼 및 파보 장염의 항체보유 실태 조사)

  • Sung, Ki-Chang;Lee, Eun-Woo;Park, Chang-Eun
    • Korean Journal of Veterinary Service
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    • v.33 no.3
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    • pp.213-221
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    • 2010
  • The results from a total of 412 blood samples consisted of 187 samples from regular visiting group (RV), 94 samples from first visiting group (FV), 52 samples from abandoned group (A), 54 samples from special breeder group (SB), and 25 samples from preliminary breeder group (PB) showed that RV(94.7%) and SB(88.9%) groups had the higher levels of protective antibody, but PB (36.0%) group revealed the lowest level. Among 96 blood samples with lower protective antibody levels, 14 samples (14.6%), 72 samples (75.0%) and 10 samples (10.4%) were below the protective antibody levels to distemper/parvo-virus, distemper only and parvovirus only, respectively. These results implied that antibody to parvovirus was well generated than that to distemper. Eighty six samples (20.9%) showed the protective antibody titer under 1:96 to distemper and 24 samples (5.8%), the protective antibody titer under 1:40 to parvovirus.

END SN Antibody Titers of Sows and Piglets Vaccinated with Living HC Vaccine During '83 Hog Cholera Outbreaks in Korea ('83 돈(豚)콜레라 유행시(流行時)의 면역모돈(免疫母豚)과 자돈(仔豚)의 END혈청중화항체가(血淸中和抗體價) 조사(調査))

  • Jeon, Yun-seong;Yeh, Jae-gil;Seo, Ik-soo
    • Korean Journal of Veterinary Research
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    • v.25 no.1
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    • pp.69-75
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    • 1985
  • Hog cholera serum neutralizing antibody of piglets and sows were titrated by means of END SN method. The piglets of a variety of ages, precolostrally immunized with LOM living HC vaccine were subjected to the test. The sows were vaccinated with lapinized living HC vaccine after 25 days from the parturition. Throughout the studies the following results were obtained and summarised. 1. Hog cholera antibody titers of inbred sows immunized with lapinized living HC vaccine after 25 days from parturition were high except Hampshire group(Table 2). 2. Sows, different stage of the pregnancy or the day of parturition, and of 3 way crossed, that were immunized with lapinized living HC vaccine have shown no significant difference on HC antibody titer(Table 2, 3). 3. HC antibody titers of piglets, immunized with a single dose of LOM HC vaccine before feeding colostrum, were high in case of the younger group(1 week) compare to the older(7 week) (Table 4). 4. The piglets that were booster immunized with LOM HC vaccine at the age of 7 weeks have shown an inconsistent antibody titers(Table 5).

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Production and characterization of monoclonal antibody against bovine coronavirus (소 코로나바이러스에 대한 단크론항체 생산과 특성)

  • Ahn, Jae-moon;Kang, Shien-young
    • Korean Journal of Veterinary Research
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    • v.38 no.3
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    • pp.581-588
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    • 1998
  • Eight monoclonal antibodies(MAbs) against bovine coronavirus(BCV) were produced and characterized. Three MAbs(1G9, 4H12, 5C1) specific to the S glycoprotein and two HE glycoprotein-specific MAbs(2A5, 5G4) were found to neutralize the BCV in fluorescence focus neutralization(FFN) test. Two HE-specific MAbs from the neutralizing MAbs inhibited the hemagglutinating activity of the BCV. None of the N protein-specific MAbs(1C1, 5A12, 6H1) neutralized the virus infectivity. Bovine coronavirus and mouse hepatitis virus, which belong to group II coronaviruses, were differentiated from other groups of coronaviruses(porcine transmissible gastroenteritis virus, porcine epidemic diarrhea virus, canine coronavirus) by all MAbs in fluorescence antibody test(FA), but not in FFN test.

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Comparative Diagnostic Studies on Serologic and Molecular Biological Tests Against Haemorrhagic Fever with Renal Syndrome (신증후출혈열 환자의 혈청학적 및 분자생물학적 진단 검사법 비교)

  • 우영대;문희주;배형준
    • Biomedical Science Letters
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    • v.6 no.2
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    • pp.141-149
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    • 2000
  • The etiologic agents of haemorrhagic fever with renal syndrome (HFRS) in Korea are Hantaan and Seoul virus in the genus Hantavirus, family Bunyaviridae. Antibody titers of sera from HFRS patients against Hantaan virus were measured by immunofluorescent antibody technique (IFAT), enzyme-linked immunosorbent assay (ELISA), high density composite particle agglutination (HDPA) and plaque reduction neutralization test (PRNI). PRNT and nested reverse transcriptase polymerase chain reaction (nested RT-PCR) was used for serotypic differentiation of Hantaviruses against Hantaan and Seoul virus. Eight doubtful HFRS patients showed higher fluorescent, IgG ELISA, agglutination and neutralizing antibody titer by IFAT, ELISA IgG, HDPA and PRNT, respectively Five out of them showed high IgM antibody titer by IgM capture ELISA against Hantaan virus, remarkably. Fifteen HFRS patients showed higher fluorescent antibody titer by IFAT. In PRNT, 12 out of them showed high neutralizing antibody titer against HTNV, 2 against SEOV and 1 against both viruses. In nested RT-PCR using serotype specific-primer, 3 out of them showed positive against HTNV and 1 against SEOV.

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Expression of VP2 of Aquatic Birnavirus GC-1 Isolated from Rockfish (Sebastes schlegeli), Rearing in Seawater in Korea (국내 해산양식어 조피볼락에서 분리된 수생버나바이러스 GC-1의 VP2 발현)

  • Joh, Seong-joon;Sung, Haan-woo;Lee, Yun-jeong;Kim, Jae-hong;Kang, Shien-young
    • Korean Journal of Veterinary Research
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    • v.43 no.3
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    • pp.449-456
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    • 2003
  • The VP2 gene of aquatic birnavirus, Korean isolate (GC-1) was cloned and expressed using the baculovirus expression system. The VP2 gene and VP2 partial gene, which contained a neutralizing epitope, were constructed for recombinant transfer vectors, for baculovirus expression. The expressed recombinant proteins were confirmed by indirect immuno fluorescence antibody (IFA), SDS-PAGE and Western blot. The level of expression was checked at regular time using IFA and Western blot. To measure the neutralizing activity of recombinant proteins against GC-1 strain, the antisera against recombinant proteins were produced by using guinea pigs. The result showed that the antisera neutralized the GC-1 strain. However, the neutralizing titer was higher in antisera against the VP2 gene expressed recombinant protein than that of VP2 partial gene recombinant protein.

Studies on the production and characterization of monoclonal antibodies against bovine rotaviruses isolated in Korea (소 로타바이러스(국내분리주)에 대한 단크론항체 생산 및 특성에 관한 연구)

  • Ahn, Jae-moon;Cho, Sun-hee;Kang, Shien-young
    • Korean Journal of Veterinary Research
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    • v.36 no.2
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    • pp.395-403
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    • 1996
  • Monoclonal antibodies(MAbs) against field isolates of the bovine rotavirus A strain(G6), V strain(G10) and reference I-801 strain(G8) were produced and characterized. Six MAbs(4C2, 4D9, 5E1, 5E7, 5D5, 3E4) against A strain had neutralizing activity and reacted only with the G6 bovine rotaviruses determined by fluorescence focus neutralization (FFN) test. Otherwise, five neutralizing MAbs(1G2, 2G6, 5E2, 5E12, 5H7) against I-801 strain neutralized the G6 and G8 bovine rotaviruses. Five non-neutralizing MAbs(5F12, 7F12, 5E11, 2A11, 2B12) were VP6-specific and cross-reacted with all bovine and porcine rotaviruses examined by fluorescence antibody(FA) test. None of the MAbs reacted with bovie viral diarrhea virus(BVDV) and bovine coronavirus(BCV) determined by FA and FFN test.

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Sequencing and Baculovirus-Based Expression of the Glycoprotein B2 Gene of HSV-2 (G)

  • Uh, Hong-Sun;Park, Jong-Kuk;Kang, Hyun;Kim, Soo-Young;Lee, Hyung-Hoan
    • Journal of Microbiology and Biotechnology
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    • v.11 no.3
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    • pp.482-490
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    • 2001
  • The gene for glycoprotein B (gB2) of HSV-2-strain G was subcloned, sequenced, recombinated into the lacZ-HcNPV, expressed in insect cells, and compared with the homologous gene of other HSV-2 strains. The ORF of the gB2 gene was 2,715 bp. The overall nucleotide sequence homology of te gB2 gene compared ith that of the two previously reported HSV-2 strains appeared to be over 98%. A recombinant virus named Baculo-gB2 protein in insect cells. The recombination was confirmed by a PCR and the expression was demonstrated by radio immunoprecipitation. Insect cells infected with the Baculo-gB2 virus synthesized and processed gB2 with approximately 120 kDa in the cells, and then secreted it into the culture media, where it reacted with a nomoclonal antibody to gB2. The gB2 polypeptide contained two main hydrophobic regions (a signal sequence from 1 to 23 amino acid residues, and a membrane anchor sequence from aa 745 to 798), eight N-glycosylation sites evenly distributed, and was rich in alanine (11.2%). Antibodies to this recombinant protein that were raised in mice recognized the viral gB2 and neutralized the infectivity of the HSV-2 in vitro. There results show that the gB2 protein was successfully porduced in insect cells and could be used to raise a protective neutralizing antibody. Accordingly, this particular recombinant protein may be useful in the development of a subunit vaccine.

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Different immunological features of two genetically distinct type 2 porcine reproductive and respiratory syndrome (PRRS) viruses

  • Shabir, Nadeem;Khatun, Amina;Kim, Won-Il
    • Korean Journal of Veterinary Service
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    • v.37 no.1
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    • pp.1-9
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    • 2014
  • Although it has been generally accepted that porcine reproductive and respiratory syndrome virus (PRRSV) induces weak and delayed protective immunity after infection, it is unclear that the same immunological features can be applicable to all PRRS viruses because huge genetic variation exists even among the same genotypes of PRRSV (Type 1 and 2). In the current study, two genetically distinct type 2 PRRSV strains (VR-2332 and JA142) which showed approximately 90% nucleotide homology based on ORF5 sequences were characterized by both in vitro and in vivo assessments to determine the immunological features of the viruses. For in vitro assessment, porcine alveolar macrophages (PAM) were infected with the viruses at $10^{-3}$ multiplicity of infection (MOI) and then supernatants and cells were collected separately at 36 hrs post infection to determine the relative expression levels of IL-$1{\alpha}$, IL-12, TNF-${\alpha}$ and INF-${\alpha}/{\beta}$ by quantitative RT-PCR. In addition, five PRRSV-free pigs were inoculated with either of JA142 or VR2332 for in vivo assessment. Serum samples were collected every week until 6 weeks post challenge. The serum samples were analyzed for the levels of viremia, PRRSV nucleocapsid-specific antibody and virus neutralizing antibody. Based on those assessments, the two viruses showed different patterns of cytokine expression in PAM and immune responses in pigs after infection. These results indicate that genetically distinct PRRSV strains have different immunological features, which might be criteria for virus classification and selection of candidate virus strains for vaccine development in the future.