• Title/Summary/Keyword: B3 antibody

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Studies on enzyme-linked immunosorbent assay(ELISA) for detection of antibody to Brucella abortus (효소면역법을 이용한 Brucella abortus 항체 검출에 관한 연구)

  • 심항섭;국정희;정봉수;고태오;조중현;박유순
    • Korean Journal of Veterinary Service
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    • v.21 no.2
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    • pp.107-115
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    • 1998
  • In order to establish a rapid, sensitive and specific diagnostic method for detection of antibody to Brucella abortus, a enzyme-linked immunosorbent assay(ELISA) was adapted. The diagnostic efficacy of the established ELISA was compared with that of the standard tube agglutination test for B abortus. 1. It was found that the optimal concentration of antigen for this ELISA was 5$\mu\textrm{g}$/ml, the optimal dilution of conjugate was 1 : 2000, and the optimal dilution of serum was 1 : 200, respectively. 2. Cut off value in this ELISA was 1,102 that was determined by mean absorbance(at 492nm) of tube agglutination test negative serum added with the triple value of the standared devation. 3. The relationship between the tube agglutination test and ELISA was showen high corresponding rate with sensitivity(96.3%) and specificity(98.1%). 4. The efficacy of the ELISA for detection of B abortus antibody was compared with tube agglutination test In brucellosis outbreak farm. The sensivity of ELSIA was higher than tube agglutination test.

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The Frequency and Distribution of Unexpected Red Cell Antibodies and Analysis of Antigen Exposure

  • Yang, Sung Sik;Yang, Hoi Joo;Park, Hyun Jun;Kwon, Seog Woon
    • Korean Journal of Clinical Laboratory Science
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    • v.45 no.4
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    • pp.164-169
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    • 2013
  • Red cell alloantibodies other than naturally occurring anti-A or anti-B are called unexpected red cell antibodies, and can be detected by performing an antibody screening. The frequency and distribution of unexpected antibody identified in Asan Medical Center were analyzed. We investigated a total of 135,238 cases of antibody screening test in AMC for 3 years from 2010 to 2012. Using column agglutination techniques, antibody identification tests were performed for the cases with positive antibody screening. Among 135,238 cases, 854 (0.6%) cases showed positive results of antibody screening test. In the order of frequency, 284 (33.3%) anti-Rh, 89 (10.4%) anti-MNS, 62 (7.3%) anti-Lewis, 34 (4.0%) anti-Kidd, 10 (1.2%) anti-Duffy, and 9 (1.1%) anti-P were identified. Multiple antibodies were detected in 199 (23.3%) cases. Among 381 subjects investigated for transfusion history, 299 (78.5%) had history of transfusion while 82 (21.5%) had unknown history. Thus the incidence of unexpected antibody was higher in the group with history of transfusion than the group without (p<0.001). Also, among 435 subjects investigated for the history of pregnancy, 46 (10.6%) had no history while 389 (89.4%) had history of pregnancy, showing higher incidence of unexpected antibody in the group with history of pregnancy than the group without pregnancy (p<0.001). Evaluated amounts and frequency of antigen exposure due to transfusion and pregnancy is suggested to increase the frequency of identification of unexpected antibody.

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Development of a Novel, Anti-idiotypic Monoclonal Anti-prolactin Antibody That Mimics the Physiological Functions of Prolactin

  • Wang, Meng;Zhang, Dian-Cai;Wang, Shen-Tian;Li, Ming-Long
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.4
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    • pp.571-579
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    • 2016
  • In this work, we prepared a panel of monoclonal anti-idiotypic antibodies to ovine prolactin (oPRL) by the hybridoma technique. Among these antibodies, one anti-idotypic antibody (designated B7) was chosen for further characterization by a series of experiments. We first demonstrated that B7 behaved as a typical $Ab2{\beta}$ based on a series of enzyme-linked immunosorbent assays. Subsequently, the results of a competitive receptor-binding assay confirmed that B7 could specifically bind to the prolactin receptor (PRLR) expressed on target cells. Finally, we examined its biological activities in CHO-PRLR and Nb2 cells and observed that B7 could activate Janus kinase 2-signal transducer and activator of transcription signalling in CHO-PRLR and Nb2 cells and induce BaF3 proliferation. The present study suggests that i) B7 can serve as a PRLR agonist or PRL mimic and has potential applications in regulating mammary gland development, milk production and maintenance of lactation in domestic animals and ii) B7 may be a biological reagent that can be used to explore the mechanism of PRLR-mediated intracellular signalling.

The Role of B Cells in Transplantation and Immunopathic Diseases

  • Basten, A.
    • IMMUNE NETWORK
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    • v.10 no.3
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    • pp.81-84
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    • 2010
  • B cells, by virtue of their diverse roles in immune responses to foreign and self antigens, have become of increasing interest to the clinician as well as the basic immunologist. In particular, it is now apparent that the development of B cell unresponsiveness in antibody and T cell mediated autoimmune disorders and the transplant setting is both worthwhile and achievable.

Asymmetries in Chickens from Lines Selected and Relaxed for High or Low Antibody Titers to Sheep Red Blood Cells

  • Tu, Yunjie;Siegel, P.B.
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.3
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    • pp.323-327
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    • 2015
  • Wattle length, width, and area were measured to classify bilateral asymmetries in four lines of chickens. The lines were the S26 generation of White Leghorns selected for high (HAS) or low (LAS) response to sheep red blood cells and sublines in which selection had been relaxed for three generations (high antibody relaxed [HAR] and low antibody relaxed [LAR]). Antibody titers (AB) were greater for HAS than for HAR with both greater than for LAS and LAR which while different for males did not differ for females. The low antibody lines were heavier and reached sexual maturity at younger age than the high antibody lines. In general, wattle length, width, and area were greater in the low than high antibody lines. In 24 comparisons for bilaterality 18 exhibited fluctuating asymmetry and 6 exhibited directional asymmetry with 5 of the 6 being for wattle length. There was not a clear pattern for changes in degree of asymmetry when selection was relaxed for 3 generations. For females, the relative asymmetry (RA) of wattle area was larger ($p{\leq}0.05$) for HAR than for LAR and not different from the selected lines and relaxed lines. There were no differences among lines for RA of wattle length and width of females and wattle length, width, and area of males.

Influence of Temperature Shifts on Antibody Synthesis in the Oliver Flounder (Paralichthys olivaceus) Immunised with Formalin Killed Edwardsiella tarda Antigen

  • Jeong, Hyun-Do;Yoon, So-Hye;Jeong, Jun-Gi;Jun, Lyn-Jin;Jeong, Joon-Bum;Lee, June-Woo
    • Journal of fish pathology
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    • v.19 no.3
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    • pp.235-241
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    • 2006
  • The effects of various temperature shifts on the kinetics of the humoral antibody response in oliver flounder, Paralichthys olivaceus, immunised with formalin-killed Edwardsiella tarda, were determined by measuring the antibody production in vivo and in vitro. When fish acclimated to a high temperature and immunised at that temperature were transferred to a lower temperature (22℃ to 12℃) at a various times after immunisation, the fish showed a weaker immune response than that achieved when the fish were kept at a high environmental temperature. However, in the converse experiment (12℃ to 22℃), the magnitude of the humoral immune response was recovered independent of the time of the transfer after immunisation at low temperature, even though the peak levels of each transferred group did not reach the level found in the positive control group that was maintained and immunised at a high environmental temperature. Hence, these studies provide some evidence that the potential for antibody production in B cells of oliver flounder immunized at high temperature is not impaired by subsequent exposure to low temperature.

Characterization of KI-24, a Novel Murine Monoclonal Antibody with Specific Reactivity for the Human Immunodeficiency Virus-1 p24 Protein

  • Shin, Song-Yub;Park, Jung-Hyun;Lee, Myung-Kyu;Jang, So-Youn;Hahm, Kyung-Soo
    • BMB Reports
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    • v.33 no.1
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    • pp.92-95
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    • 2000
  • The HIV-1 p24(202-221) sequence ETINNEEEWDRVHPV HAGP contains a B-cell epitope with the earliest immune response and the highest antibody titer against anti-mouse sera obtained by immunization with p24 antigens. A novel mouse monoclonal antibody (mAb) was generated against the immunodominant B-cell epitope of the HIV-1 p24 capsid protein, p24(202-221). BALB/c mice were immunized with the four branched multiple antigenic peptide (MAP) containing the HIV-1p24(202-221) sequence, and antibody-secreting hybridoma were produced by fusion of mouse splenocytes with P3X63Ag8.653, mouse myeloma cells. One clone which produced the antigen-specific mAb named KI-24 (Isotype IgG1, light chain: ${\kappa}$) was identified. mAb KI-24 was highly specific for both the p24(202-221) and p24 proteins when analyzed by ELISA and Western blotting. Since p24(202-221) also contains a cytotoxic T-lymphocyte epitope, this specfic peptide epitope and the monoclonal antibody with specific reactivity against the p24 protein and p24(202-221) can be used in peptide vaccine development and p24 antigen detection from HIV patients.

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A comparative study on immunogenicity of the porcine epidemic diarrhea virus live-vaccine and inactivated-vaccine (돼지유행성설사병(PEDV) 생독과 사독백신의 면역형성 비교연구)

  • Kwon, Mee-Soon;Cho, Hyun-Ung;Lee, Eun-Mi;Lee, Ji-Yoog;Seo, Heyng-Seok;Im, Jeong-Cheol;Hur, Boo-Hong
    • Korean Journal of Veterinary Service
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    • v.32 no.3
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    • pp.201-207
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    • 2009
  • Porcine transmissible gastroenteritis virus (TGEV), porcine epidemic diarrhea virus (PEDV) and rotaviruses are considered as the most important causative agents of diarrhea in piglets. The study established 3 method vaccination programs to prevent PEDV. A (LL)group inoculated twice vaccinations on 2-weeks-interval during the late term of pregnant sows with PEDV live vaccine. The B (LKK) group was applied that one time single PEDV live vaccine at the pre-mate followed by the TGEV PEDV combined inactivated vaccine (twice vaccination on 2-weeks interval at the third-trimester). C (KK) group was applied to sow which inoculated twice vaccination on 2-weeks-interval during the late term of pregnant sows with by the TGEV, PEDV combined inactivated vaccine. As the result of SN test on sows in the pig farm before vaccination, antibody titers was showed 9/45 (20.0%). By comparison with the serum neutralizing antibody titers against PEDV of the vaccination programs after PEDV of the vaccination, A group and B group vaccination method was higher than those of C group in sows. In the piglets up to 2 weeks of age, A group was showed antibody titers of 17/22 (81.8%) that showed 2-128, and B group was showed antibody titers of 30/37 (81.1%) that showed 2-512, and C group was showed antibody titers of 14/28 (50.0%) that showed 2-32. On the other hand, PEDV antibody titers were tested for the survey. As the results of SN test, Aujeszky's disease survey in 54 pig farms from november 2005 to august 2006, antibody titers of 47/286 (16.4%) showed above 2. Five breeding farms were antibody titers of 38/77 (49.4%), Wanggung zone farms antibody titers of 59/85 (69.4%). In pigs farms vaccinated the first of twice PEDV live vaccine, and after 6 month, the second of twice TGEV PEDV combined inactivated vaccine (LLKK, 256-1024 titer) method was higher than those of vaccinated twice the early term of pregnant, and twice the late term of pregnant sows of PEDV live vaccine (LLLL, 32 titer).

Fine Mutational Analysis of 2B8 and 3H7 Tag Epitopes with Corresponding Specific Monoclonal Antibodies

  • Kim, Tae-Lim;Cho, Man-Ho;Sangsawang, Kanidta;Bhoo, Seong Hee
    • Molecules and Cells
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    • v.39 no.6
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    • pp.460-467
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    • 2016
  • Bacteriophytochromes are phytochrome-like light-sensing photoreceptors that use biliverdin as a chromophore. To study the biochemical properties of the Deinococcus radiodurans bacteriophytochrome (DrBphP) protein, two anti-DrBphP mouse monoclonal antibodies (2B8 and 3H7) were generated. Their specific epitopes were identified in our previous report. We present here fine epitope mapping of these two antibodies by using truncation and substitution of original epitope sequences in order to identify minimized epitope peptides. The previously reported original epitope sequences for 2B8 and 3H7 were truncated from both sides. Our analysis showed that the minimal peptide sequence lengths for 2B8 and 3H7 antibodies were nine amino acids (RDPLPFFPP) and six amino acids (PGEIEE), respectively. We further characterized these peptides in order to investigate their reactivity after single deletion and single substitution of the original peptides. We found that single-substituted 2B8 epitope (RDPLPAFPP) and dual-substituted 3H7 epitope (PGEIAD) showed significantly increased reactivity. These two antibodies with high reactivity for the short modified peptide sequences are valueble for developing new peptide tags for protein research.

Generation of Isotype Switch Variants form Hybridoma cells Producing anti-Streptococcus penumoniae 6B Polysaccharide Antibody

  • Kim, Jihye;Eunja Ryu;Park, Moon-Kook
    • Journal of Microbiology
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    • v.37 no.3
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    • pp.180-184
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    • 1999
  • hybridoma cells producing IgM anti-pneuococcal 6B polysaccharide antibodies were induced to switch to IgG-producing cells in vitro by treating with acridine orange. Treating 0.5 $\mu\textrm{g}$/ml of acridine orange for 24 hours generated maximal number of variant cells. The maximal isotype switch frequency was 3${\times}$10-5, which is about 30-fold higher than the frequency of spontaneous switching. Resulting IgG-producing variants were enriched by sib selection and ELISA spot assay. Two IgG3-producing variant cells were finally cloned by limiting dilution. The variant cells produced similar amounts of antibodies as their parental cells did. The two switched antibodies had similar reactivity to pneumococcal 6B polysaccharide. When compared to their parental IgM antibodies, the switched IgG3 than that of IgM antibody. The antibodies will be useful as essential tools for comparative study of the role of heavy chain isotypes in protection against Streptococcus pneumoniae.

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