• Title/Summary/Keyword: B3 antibody

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A Divalent Immunotoxin Formed by the Disulfide Bond between Hinge Regions of Fab Domain

  • Choe, Seong Hyeok;Kim, Ji Eun;Lee, Yong Chan;Jang, Yeong Ju;Choe, Mu Hyeon
    • Bulletin of the Korean Chemical Society
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    • v.22 no.12
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    • pp.1361-1365
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    • 2001
  • Recombinant immunotoxins are hybrid cytotoxic proteins designed to selectively kill cancer cells. A divalent immunotoxins, [B3(FabH1)-PE38]2, was constructed by recombining Fab domain of B3 antibody as a cell-targeting domain and Pseudo monas exotoxin A (PE) as a cytotoxic domain. Monoclonal antibody, B3, is the murine antibody (IgG1k) directed against Lewis Y-related carbohydrate antigens, which are abundant on the surface of many carcinomas. Fab fragment of this antibody was used in this study with the modified hinge sequence where last two cysteines out of three were mutated to serine. PE is a 66 kDa bacterial toxin that kills eukaryotic cells by inhibiting protein synthesis with ADP ribosylation of ribosomal elongation factor 2 (EF2). Fc region of B3 antibody was substituted with the truncated form of PE (38 kDa, PE38) on DNA level. [B3(FabH1)-PE38]2 was formed by disulfide bond between cysteines in the modified hinge region of B3(FabH1)-PE38. Each polypeptide for recombinant immunotoxins was overexpressed in Escherichia coli and collected as inclusion bodies. Each inclusion body was solubilized and refolded, and cytotoxic effects were measured. Divalent immunotoxins, [B3(FabH1)-PE38]2, had ID50 values of about 10 ng/mL on A431 cell lines and about 4 ng/mL on CRL1739 cell lines. Control immunotoxins, B3(scFv)-PE40, had ID50 values of about 28 ng/mL on A431 cell lines and about 41 ng/mL on CRL1739 cell lines. Divalent immunotoxins, [B3(FabH1)-PE38]2, had higher cytotoxic effects than B3(scFv)-PE40 control immunotoxins.

A Role of Neutrophils in Anti-Candida Monoclonal Antibody Protection Against Vaginal Infection due to Candida albicans (캔디다질염에 효과가 있는 단항체에 대한 호중구의 역할)

  • 한용문
    • YAKHAK HOEJI
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    • v.47 no.3
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    • pp.190-194
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    • 2003
  • As previously reported, an immunoglobulin M (IgM) monoclonal antibody (MAb) B6.1, specific for a cell wall B-l,2-mannotriose, was protective against vaginal infection due to Candida albicans when mice were treated with the antibody. In this study, the role of neutrophil was examined in the protective effect of MAb B6.1 against vaginal infection. To deplete neutrophils, mice were given intravenously rat anti-mouse neutrophile MAb RB6-8C5 prior to intraperitoneal administration of MAb B6.1 to these mice. The mice were examined for antibody in their reproductive tract. By an ELISA, MAb B6.1 was found in the vaginal homogenates, but no antibody was detected in vaginal lavage materials. The neutropenia was induced by a single dose of the anti-neutrophil antibody, but lymphocytes were also partially depleted. The protective effect of MAb B6.1 was decreased when mice pretreated with MAb RB6-8C5 were given the anti-Candida antibody before challenge with C. albicans yeast cells intravaginally. These results show that neutrophils are involved in the MAb B6.1 protection against Candida vaginal infection.

Changes in SARS-CoV-2 antibody titers 6 months after the booster dose of BNT162b2 COVID-19 vaccine among health care workers

  • Takeshi Mochizuki;Takaki Hori;Koichiro Yano;Katsunori Ikari;Ken Okazaki
    • Clinical and Experimental Vaccine Research
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    • v.12 no.2
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    • pp.116-120
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    • 2023
  • Purpose: In Japan, the data on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibody titers after the booster dose of the coronavirus disease 2019 (COVID-19) vaccine are insufficient. The aim of this study is to evaluate changes in SARS-CoV-2 antibody titers before, 1, 3, and 6 months after the booster dose of the BNT162b2 COVID-19 vaccine among health care workers. Materials and Methods: A total of 268 participants who received the booster dose of the BNT162b2 vaccine were analyzed. SARS-CoV-2 antibody titers were measured before (baseline) and at 1, 3, and 6 months after the booster dose. Factors associated with changes in SARS-CoV-2 antibody titers at 1, 3, and 6 months were analyzed. Cutoff values at baseline were calculated to prevent infection of the omicron variant of COVID-19. Results: The SARS-CoV-2 antibody titers at baseline, and 1, 3, and 6 months were 1,018.3 AU/mL, 21,396.5 AU/mL, 13,704.6 AU/mL, and 8,155.6 AU/mL, respectively. Factors associated with changes in SARS-CoV-2 antibody titers at 1 month were age and SARS-CoV-2 antibody titers at baseline, whereas changes in SARS-CoV-2 antibody titers at 3 and 6 months were associated with the SARS-CoV-2 antibody titers at 1 month. The cutoff values of the SARS-CoV-2 antibody titers at baseline were 515.4 AU/mL and 13,602.7 AU/mL at baseline and 1 month after the booster dose, respectively. Conclusion: This study showed that SARS-CoV-2 antibody titers increase rapidly at 1 month after the booster dose of the BNT162b2 vaccine and begin to decrease from 1 to 6 months. Hence, another booster may be needed as soon as possible to prevent infection.

Screening of the Antigen Epitopes of Basic Fibroblast Growth Factor by Phage Display

  • Xiang, Junjian;Zhong, Zhenyu;Deng, Ning;Zhong, Zhendong;Yang, Hongyu
    • BMB Reports
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    • v.38 no.3
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    • pp.290-293
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    • 2005
  • In order to investigate the epitope of basic fibroblast growth factor (bFGF) and its immunogenicity, the epitopes of bFGF were screened from the phage display library with monoclonal antibody GF22, which can neutralize the bio-activity of bFGF. By three rounds of screening, the positive phage clones with bFGF epitopes were selected, which can effectively block the bFGF to bind with GF22. Sequence analysis showed that the epitopes shared a highly conservative sequence (Leu-Pro-Pro/Leu-Gly-His-Phe/Ile-Lys). The sequence of PPGHFK was located at 22-27 of the bFGF. The specific immuno-response of mouse could be highly induced by phage clones with the epitopes. And the anti-bFGF activity induced by LPGHFK was 3 times higher than the original sequence, which showed that the mimetic peptide LPLGHIK might be used as a tumor vaccine in the prevention and treatment of tumor.

Affinity Maturation of an Anti-Hepatitis B Virus PreS1 Humanized Antibody by Phage Display

  • Yang, Gi-Hyeok;Yoon, Sun-Ok;Jang, Myung-Hee;Hong, Hyo-Jeong
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.528-533
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    • 2007
  • In a previous study we generated an anti-Hepatitis B Virus (HBV) preS1 humanized antibody (HzKR127) that showed in vivo HBV-neutralizing activity in chimpanzees. However, the antigen-binding affinity of the humanized antibody may not be sufficient for clinical use and thus affinity maturation is required for better therapeutic efficacy. In this study, phage display technique was employed to increase the affinity of HzKR127. All six amino acid residues (Glu95-Tyr96-Asp97-Glu98-Ala99-Tyr100) in the heavy (H) chain complementary-determining region 3 (HCDR3) of HzKR127 were randomized and phage-displayed single chain Fv (scFv) library was constructed. After three rounds of panning, 12 different clones exhibiting higher antigen-binding activity than the wild type ScFv were selected and their antigen-binding specificity for the preS1 confirmed. Subsequently, five ScFv clones were converted to whole IgG and subjected to affinity determination. The results showed that two clones (B3 and A19) exhibited an approximately 6 fold higher affinities than that of HzKR127. The affinity-matured humanized antibodies may be useful in anti-HBV immunotherapy.

Frequencies and Distributions of Unexpected Antibodies at a General Hospital in the Daejeon of Korea (대전지역 일개 종합병원에서의 비예기 항체 발생현황과 분포)

  • Kim, Jae-Jung
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.3
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    • pp.354-358
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    • 2018
  • When preparing for a blood transfusion, the presence and types of unexpected antibodies should be identified through screening tests. Using the DiaMed-ID system, antibody identification among unexpected antibody screening tests performed at a university hospital in Daejeon area for two years from January 2016 to December 2017 were limited to 55 patients and were predominantly women. A total of 36 patients (65.5%) belonged to the Rh group, 7 (12.7%) patients to the Lewis group, 4 (7.3%) patients to the Kidd and Duffy groups, 3 patients (5.5%) to the MNS group, and 1 (1.8%) to the Rh+Kidd combined group. In the Rh group, 19 (34.5%) patients had Anti-E single antibody, 5 (9.1%) patients had Anti-D single antibody, 4 (7.3%) patients had Anti-E/-c, 4 (7.3%) patients had Anti-C/-e, and 1 (1.8%) patient had $Anti-E/-c/-Jk^b$. In the Lewis group, three (5.5%) patients had both $Anti-Le^a$ and $Anti-Le^b$. In the Kidd group, one (1.8%) patient had $Anti-Jk^a$ and three (5.5%) patients had $Anti-Jk^b$. In the Duffy and MNS groups, only single antibody was found: one (1.8%) patient with $Anti-Fy^a$, three (5.5%) patients with $Anti-Fy^b$, two (3.6%) patients with Anti-M, and one (1.8%) patient with Anti-S. This study reflects the recent frequencies and distributions of unexpected antibodies in Daejeon, which would be helpful for the efficient preparation for transfusions.

Epidemiological studies on infectious bursal disaese of chickens in southern area of kangwon province (강원 남부지역 산란계 및 육계의 전염성 F낭병에 대한 역학 조사연구)

  • 최문희;이시창;박원헌;김지태;김남선;권종규;이유섭
    • Korean Journal of Veterinary Service
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    • v.19 no.3
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    • pp.221-226
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    • 1996
  • IBD's antibody level and morphological change of immune organ was examined in chicken. The results were as follows ; 1. Seventy percent at 42 day and 40% at 45 day of age chickens were reacted positively by the agar gel immunodiffusion test and 42 day and 45 day of age chickens indicated 1859, 1425 by the ELISA test, respectively. 2. In 2 and 5 day young broiler chickens, the level of maternal antibody was not proper. B/B ratio showed low level, but S/B ratio and BS were normal. 3. In layer, 100% of 8 and 86 day of age chicken and 70% at 40 day of age chicken had antibody aganist IBDV. The level of antibody was high as 2293 and 3336 in 8 and 86 day of age chicken, while was low as 1186 in 40 day of age chicken. B/B ratio showed low level and S/B ratio high level, but BS was normal.

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Effect of 65 kDa Buffalo Placental Protein on B-Cell Proliferation and Antibody Response

  • Deepthi, K.;Rajput, Y.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.3
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    • pp.432-437
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    • 2002
  • Immunosuppressive potential of 65 kDa buffalo placental protein (bPP65) on B-cell proliferation in vitro and antibody response in vivo was evaluated. B-cell proliferation was estimated by measuring incorporation of tritiated thymidine in buffalo lymphocytes while primary antibody responses against phytohaemagglutinin (PHA) or keyhole limpet haemocyanin (KLH) were evaluated in mice. bPP65 suppressed proliferation of lipopolysaccharide (a B-cell specific mitogen)-stimulated buffalo lymphocytes in vitro indicating suppression of B-cells. This suppression was dose dependent over the protein concentration range $25-100 {\mu}g/ml$. Primary antibody responses in mice against PHA and KLH in presence of bPP65 were lower as compared to in its absence but these were not statistically significant. Amino acid composition data of bPP65 and BSA suggested that bPP65 is different from BSA.

Studies on the Enhancing Effect of Polymyxin B on the Antibodies Response of Enterobacterial Antigens (Pomyxin B의 장계세균항원(腸系細菌抗原)에 대(對)한 항체산생(抗體産生) 증강작용(增强作用)에 관(關)한 연구(硏究))

  • Lee, Jae-Koo
    • The Journal of the Korean Society for Microbiology
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    • v.6 no.1
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    • pp.29-40
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    • 1971
  • Various kinds of antibiotics are generally believed to have inhibitory effects on the antibody response. However, as polymyxin B which belongs to the cyclic polypeptide group of antibiotic was found to have some enhancing effects on the antibody response of rabbits to enterobacterial common antigen(CA) under specified conditions, experiments were carried out on this problem with the following results. 1. When mixture of polymyxin B and CA derived from Salmonella typhimurium(STM) was treated 30 minutes at $37^{\circ}C$ and injected three times into rabbits by intravenous route, the antibody response to CA was weaker than rabbits injected CA only. 2. Mixture of polymyxin B and CA showed a marked antibody production when injected into rabbits primed with small amounts of heat-extracted antigen of STM, while the injection of CA alone showed low titers of response. 3. Mixture of polymyxin B and heat-extracted CA-containing antigen of Escherichia coli 014 also showed a increased antibody production than CA alone in rabbits primed with antigen of STM. 4. The effect of polymyxin B appeared in different ways. This antibiotic did not enhance the CA antibody response in rabbits primed with small amounts of E. coli 0111 and 055, but enhance in rabbits primed with Shigella flexneri. 5. No enhancing effect on the antibody response was observed by polymyxin B in rabbits primed with CA. 6. No enhancing effect on the antibody response was also noted in rabbits primed with STM antigen in case polymyxin B and CA were administered simultaneously but in veins of different places. 7. Bacitracin did not enhance the CA antibody response in primed rabbits with STM antigen, but neomycin slightly enhance the response. 8. Lipopolysaccharide showed no priming effect on the CA antibody response, and no enhancement of the CA antibody response in rabbits printed with STM. 9. The priming effect of STM antigen against CA antibody response was very weak as compared with the effect of CA derived from STM antigen.

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Purification of Anti-HLA Antibodies in Human Placenta Sera (사람 태반혈청내의 항HLA항체 정제)

  • Lim, Byung-Uk;Han, Hoon;Rhyu, Moon-Gan;Kim, Tae-Kyu;Kim, Gum-Ryong;Lee, Chong-Hoon
    • The Journal of the Korean Society for Microbiology
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    • v.19 no.1
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    • pp.79-83
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    • 1984
  • To determine the existence of anti-HLA antibodies finally in 220 human placental extracts to be proved negative antiserum by previous anti-HLA A,B,C antibody screening procedure, the present study was performed by fractionation of immunoglobulins using saturated ammonium sulfate and by simple batch method on DEAE cellulose. Thereafter using known 150 T-lymphocyte panels, complement-dependent microlymphocytotoxicity test was performed to observe the existence of anti-HLA antibodies and the degree of the antibody response of the concentrates. The following results were obtained: 1. Of total 141 placental sera concentrated 45 cases(31.9%) were showed significant anti-HLA A,B,C antibody response after concentration(Excellent, 19(13.5%), Good, 3(2.1%), Weak, 23(16.3%)). 2. Anti-HLA specificities of placental sera obtained after concentration were A2, A24, B13, B27, B44, B51, CN1, C7. 3. A new type C new-1 anti-HLA antibody that is only expressed in Korean people, was obtained. 4. 79 placental sera purrified by simple batch method using DEAE cellulose were showed negative anti-HLA antibody responses.

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