• Title/Summary/Keyword: B3 antibody

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Studies on the development of enzyme linked immuno-sorbent assay (ELISA) for hepatitis B surface antigen (HBsAg) by monoclonal antibodies of different affinity constants

  • Kim, Gye-Won;Hong, Sung-Youl;Shin, Soon-Cheon;Lee, Sung-Hee;Kim, Won-Bae
    • Archives of Pharmacal Research
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    • v.10 no.1
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    • pp.18-24
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    • 1987
  • Mouse monocolonal antibodies to Hepatitis B surface antien (HBsAg) were prepared and their functional capabilities tested by the method of solid phase enzyme linked immuno sorbent assay (ELISA). HBsAg binding studies inicated that one monoclonal antibody 6E-1-1 bound more HBsAg at a faster rate than the other monoclonal antibodies. Also, for the binding inhibition studies with the selected monoclonal antibody 6E-1-1, one monoclonal antibody 8D-3-6 didn't exhibit binding inhibition for HBsAg. Then, a simultaneous ELISA method was developed for the immunodiagnosis of HBsAg. Different combinations of two monoclonal antibodies as solid phase and horseradish peroxidase (HRPO) labeled phase were studied. The combination of monoclonal antibody of higher affinity constant (6E-1-1) immobilized in a solid phase and monoclonal antibody of lower affinity constant (8D-3-6) as a HRPO laeled phase was more sensitive when two monoclonal antibodies of different affinity constants for HBsAg were prepared.

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Efficacy of Newcastle Disease Virus $B_1$Vaccine in Quails (메추리에서의 Newcastle균 바이러스 $B_1$백신의 효능시험)

  • 박형철;최정옥
    • Korean Journal of Poultry Science
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    • v.19 no.1
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    • pp.17-25
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    • 1992
  • The efficacy of B$_1$l live vaccine which is used successfully in chicken was examined against Newcastle disease in quails. A total of 480 male quails were divided into 4 groups, of which 3 groups were vaccinated via drinking water, eye instillation and spraying method and the remaining was employed as a nonvaccinated control group. At 3 weeks after the first vaccination a part of quails in each group was revaccinated. Efficacy of the vaccine was evaluated by the antibody responses and the protection rates after challenge with a virulent NDV. Vaccination of quails with $B_1$ NDV at 10 days of age resulted in beneficial effect compared to nonvaccinated group regardness of vaccination methods adopted although general protection rates were considerably low. Twice vaccination gave higher protection than once vaccination. Hemagglutination inhibition antibody responeses were significantly higher in groups of quails vaccinated by spray and eye instillation method than by drinking water administration. Antibody responses were marked at 2 weeks onward and until 5 to 7 weeks after vaccination. Antibody responses were rapid and marked after second vaccination. However, antibody level did not last longer than 5 to 7 weeks postvaccination. Vaccine caused no adverse effect on quails in terms of growth'rate, body temperature or clinical signs.

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Immunological Properties of Recombinant Hepatitis B Surface Antigen Expressed in Mammalian Cell(C127)

  • Lee, Young-Soo;Kim, Byong-Kak;Choi, Eung-Chil
    • Archives of Pharmacal Research
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    • v.21 no.5
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    • pp.543-548
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    • 1998
  • We examined the immunological properties of the recombinant hepatitis B surface antigen (r-HBsAg) which was expressed in mammalian cell (C127). The cross-immunity of r-HBsAg and plasma-derived hepatitis B surface antigen (p-HBsAg) were tested using Western blotting and ELISA with guinea pig polyclonal antibody and naturally infected human-derived antibody and the both antigens show the same results in their response pattern and intensity, which indicate they have a good cross-immunity. from the measurement of $ED_{50}$ after formalin- or heat-inactivation, both r-HBsAg and p-HBsAg and p-HBsAg showed $ED_{50}$ of 0.2-0.3 in formalin-inactivaton, while r-HBsAg was 0.05-0.09 and p-HBsAg was 0.03-0.07 in heat-inactivation, which means heat-inactivation method is 3-4 times superior in immunogenicity. In the immunopersistency test performed in guinea pig for the period of 3 months with two different adjuvants, antibody titer was 34.2 with muramyl dipeptide adjuvant, which was 1.8 times greater than the antibody titer of 18.9 with $AIPO_{4}$ adjuvant. the mutagenicity of r-HBsAg has the same cross-immunity with p-HBsAg, and heat-inactivation method and muramyl dipeptide adjuvant allow development of r-HBsAg vaccine with excellent immunogenicity.

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Neuromyelitis optica spectrum disorders with an inverted V sign on spinal cord magnetic resonance imaging: anti-aquaporin-4 antibody and functional vitamin B12 deficiency (척수에 뒤집힌 V징후를 가진 시신경척수염: 항아쿠아포린-4항체와 비타민 B12 기능적 결핍)

  • Sung Jo Bang;Sohyeon Kim;Young Seok Jeong;Seo Hyeon Lee;Hung Youl Seok
    • Journal of Medicine and Life Science
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    • v.19 no.3
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    • pp.130-133
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    • 2022
  • Several studies have reported a possible link between anti-aquaporin-4 antibody and vitamin B12 deficiency in neuromyelitis optica spectrum disorder (NMOSD). Bilaterally symmetric hyperintense signals on magnetic resonance imaging (MRI) of the posterior columns, called the inverted V sign, are a characteristic feature of subacute combined degeneration associated with vitamin B12 deficiency. We report a patient with anti-aquaporin-4 antibody-positive NMOSD and an inverted V sign on MRI of the spinal cord and address the association between anti-aquaporin-4 antibody and functional vitamin B12 deficiency.

Comparison of immunogenecities of three beta-nodavirus proteins, capsid protein, non-structural protein B1 and B2 in olive flounder

  • Cha, Seung-Ju;Do, Jeong-Wan;Ko, Myoung-Seok;Kim, Jin-Woo;Park, Jeong-Woo
    • Journal of fish pathology
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    • v.22 no.3
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    • pp.219-228
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    • 2009
  • The genomic and subgenomic RNAs of fish nodavirus encode the four proteins, protein A, capsid protein, non-structural protein B1 and B2. In this study, we describe the immune response of olive flounder Paralichthys olivaceus immunized with live fish nodavirus or recombinant capsid protein, non-structural protein B1 and B2 expressed in E. coli. Nodavirus-infected flounder produced antibodies to capsid protein, B1 and B2 and nodavirus-neutralizing activities were detected in the serum of the nodavirus-infected flounder. The flounder were immunized against the three recombinant proteins of fish nodavirus and the sera from these immunized fishes were assayed for nodavirus-specific antibody by ELISA and a neutralization test. In the immunized flounder, all three recombinant proteins induced the production of similar levels of antibody, but only the antibody to capsid protein significantly neutralized nodavirus. These results indicate that all three nodaviral proteins are immunogenic in flounder, but only the capsid protein can induce neutralizing antibody against nodavirus.

Periplasmic Expression of a Recombinant Antibody (MabB9) in Escherichia coli

  • Chang, Hae-Choon;Kwak, Ju-Won
    • Journal of Microbiology and Biotechnology
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    • v.7 no.5
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    • pp.299-304
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    • 1997
  • Expression in the periplasm of Escherichia coli of cloned heavy and light chain cDNAs for Fab fragment of a murine monoclonal antibody MabB9 (${\gamma}2b$, K), specific for human plasma apolipoprotein B-100 of LDL, was studied. For the purpose, a vector for two-cistronic expression of the heavy chain cDNA, at the 5' terminus, and light chain cDNA, at the 3' terminus, was constructed using the signal sequences, pelB (for heavy chain) and ompA (for light chain) in a pET vector system. The constructed vector was transformed into E. coli BL21(DE3). The expressed heavy chain (25 kDa) and light chain (23 kDa) of the antibody molecule were detected in total cell extracts as well as in the periplasmic extracts of E. coli.

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3C8, a new monoclonal antibody directed against a follicular dendritic cell line, HK

  • Lee, In Yong;Lee, Joonhee;Park, Weon Seo;Nam, Eui-Cheol;Shin, Yung Oh;Choe, Jongseon
    • IMMUNE NETWORK
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    • v.1 no.1
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    • pp.26-31
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    • 2001
  • Background : Follicular dendritic cells (FDCs) play key roles during T cell-dependent humoral immune responses by allowing antigen-specific B cells to survive, proliferate, and differentiate within the FDC networks of secondary follicles, i.e., germinal centers (GC). Methods: A novel monoclonal antibody, 3C8, was generated by immunizing with an FDC line HK, in order to understand the molecular signals involved in the FDC-B cell interactions in the microenvironment of the GC. Results: The 3C8 antibody did not bind to mononuclear cells, including T cells, B cells, and monocytes. Murine L929 and human skin fibroblasts exhibited no or little reactivity to 3C8. However, 3C8 specifically recognized HK cells by flowcytometry. Furthermore, the antigen recognized by 3C8 was restricted to the GC of the human tonsil. Dendritic networks of the GC were intensely stained by 3C8, but cells outside the GC were not. Conclusion: Our results suggest that the antigen 3C8 may play some unique role on FDCs during the GC reactions.

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Increase of Spacer Sequence Yields Higher Dimer $(Fab-Spacer-Toxin)_{2}$ Formation

  • Yoo Mee-Hyeon;Won Jae-Seon;Lee Yong-Chan;Choe Mu-Hyeon
    • Journal of Microbiology and Biotechnology
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    • v.16 no.7
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    • pp.1097-1103
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    • 2006
  • The divalent antibody-toxins are expected to have increased binding avidities to target cells because of the two cell-binding domains. However, previous studies showed that the refolding yield of divalent antibody-toxin is very low, and it is assumed that homodimer formation of antibody-toxin is strongly interfered by the repulsion between the two large toxin domains that come close to each other during dimer formation. In this study, B3 antibody was used as a model antibody, and its Fab domain was used to construct three different kinds of Fab divalent molecules, $[B3(Fab)-toxin]_{2}$. The monomer Fab-toxin molecules were made by fusing the Fab domain of monoclonal antibody B3 to PE38, a truncated mutant form of Pseudomonas exotoxin (PE), and a connecting sequence that contained spacer amino acid sequence (G4S)n (n=l, 2, 3) was inserted between Fab and PE38. The prepared divalent molecules were $[Fab-S\;1,\;2,\;3-PE38]_{2}\;(=[Fab-SKPCIST-KAS(G_{4}S)nGGPE-PE38]_{2}\;(n=1,\;2,\;3))$, and they are derivatives of previously studied $[Fab-H2cys-PE38]_{2}\;(=[Fab-SKPCIST-KASGGPE-PE38]_{2})$. In $[Fab-Sl,\;2,\;3-PE38]_{2}$, two Fab-S1, 2, 3-PE38 monomers were covalently linked by the disulfide bond bridge made from cysteine in the -SKPCIST- sequence. The insertion of spacer amino acids after the disulfide bridge resulted in a 12-18 fold higher yield of dimer formation than previously constructed $[Fab-Hlcys-PZ38]_{2}[7]$, 3-4-fold higher than $[Fab-ext-PZ38]_{2}[25]$. These two molecules have less amino acid spacer sequence between the disulfide bridge and PE38 domain. The design of $[Fab-PE38]_{2}$ in this study gave molecules with a higher refolding yield. The results of cytotoxicity assay showed a higher cytotoxic effect of these divalent molecules than that of the monovalent scFv-PE38 molecule.

Distribution of Antibodies Against Hepatitis B virus and Hepatitis C virus in Human Sera (사람 혈청내 B형과 C형 간염바이러스에 대한 항체분포)

  • Choi, Jin-Sub;Seong, In-Wha
    • The Journal of Korean Society of Virology
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    • v.27 no.2
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    • pp.129-135
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    • 1997
  • Serum samples from 123 males and 123 females collected by age in 1996 were analyzed for antibodies against surface antigen of Hepatitis B virus and C22-3, C200 antigens of Hepatitis C virus. Sera from the children under the age of 10 showed 30% seropositivity to the surface antigen of Hepatitis B virus, 33.3% in $10{\sim}19$ year group, 20% in $20{\sim}29$ year group, 17.6% in $30{\sim}39$ year group, 3.3% in $40{\sim}49$ year group, 5.9% in $50{\sim}59$ year group, 8,3% in $60{\sim}69$ year group, 2.9% in $70{\sim}79$ year group, but antibody could not found in $80{\sim}86$ year group. 12 out of 123 male sera were positive, 19 out of 123 female sera were positive and overall rate of positivity of antibody against surface antigen of Hepatitis B virus was 12.6%. Serum samples from peoples under the age of 30 had not antibody against C22-3, C200 antigens of Hepatitis C virus. The positivity rate was 2.9% in $30{\sim}39$ year group. 5 out of 30 sera from $40{\sim}49$ year age group were positive, and 3 positive sera showed extremely high titer (1:524,288) but the titers of two remaining sera were 1:32, 1:8,192 respectively. 5.9% was positive in $50{\sim}59$ year group, 8.3% in $60{\sim}69$ year group, 11.8% in $70{\sim}79$ year group but all negative in $80{\sim}86$ year group 6 out of 123 male sera were positive (4.9%), 9 out of 123 female sera were positive (7.3%). Overall rate of positivity of antibody against C22-3, C200 antigen of Hepatitis C virus was 6.1 %. None out of 246 sera had both antibodies against Hepatitis B virus and Hepatitis C virus.

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Monoclonal Antibody against leucocyte CD11b(MAb 1B6) increase the early mortality rate in Spraque Dawley with E. coli pneumonia (백혈구 CD11b에 대한 단 클론 항체 (MAb 1B6)는 Spraque Dawley의 E. coli 폐렴의 조기 사망률을 증가시킨다)

  • Kim, Hyung Jung;Kim, Sung Kyu;Lee, Won Young
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.4
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    • pp.579-589
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    • 1996
  • Background : Activation of neutrophil is critical for the clearance of microorganisms and toxic host mediators during sepsis. Unfortunately the activated neutrophil and its toxic byproducts can produce tissue injury and organ dysfunction. The leucocyte CD11/18 adhesion complex regulates neutrophil-endothelial cell adhesion, the first step in neutrophil migration to sites of injection and inflammation. To investigate the potential of neutrophil inhibition as a treatment strategy for sepsis, we evaluated the effects of monoclonal antibody against CD11b (MAb 1B6) in rats intrabronchial challenged with Escherichia coli. Methods : Animals were randomly assigned to receive monoclonal antibody against CD11b (1 mg/kg, sc) and bovine serum albumin(BSA, 1 mg/kg, sc) 6 hr before, at 0 and 6 hr after intrabronchial challenge of $20x10^9$ CFU/kg E. coli 0111. Animals were randomized to treat either 24, 60 or 90% oxygen after bacterial challenge and begining 4 hr after inoculation, all animals were received 100 mg/kg ceftriaxone qd for 3 days. Peripheral and alveolar neutrophil(by bronchoalveolar lavage) counts and lung injury parameters such as alveolar-arte rial $PO_2$ difference, wet to dry lung weight ratio and protein concentration of alveolar fluid were measured in survived rats at 12 hr and 96 hr. Results : Monoclonal antibody against CD11b decreased circulating and alveolar neutrophil especially more in 12 hr than in 96 hr The lung injury parameters of antibody-treated animals were not different from those of BSA-treated animals. but It was meaningless due to small number of survived animals. The early(6 hr) mortality rate was significantly increased in antibody-treated group(51%) compared to BSA-treated group(31%) (P=0.02) but late(from 12 hr to 72 hr) mortality rate was not different in antibody-treated group(44%) from BSA-treated group(36%) (P =0.089). Conclusion : Leucocyte CD11b/18 adhesion molecule is known to regulate neutrophil migration to the site of infection and inflammation. The monoclonal antibody against CD11b decreased alveolar neutrophil in rats with pulmonary sepsis and increased early mortality rate. Therefore, we can speculate that monoclonal antibody against CD11b blocks of alveolar recruitment of neutrophils, impairs host defense mechanism and increases early mortality rate of pulmonary sepsis in rat.

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