• Title/Summary/Keyword: hepatitis B surface antigen(HBsAg)

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Effects of Glucose and Glutamine Concentrations on Hybridoma Growth ann Anti-HBsAg MAb Production in Enriched Medium (농축배지에서 Glucose와 Glutamine 농도가 하이브리도마 증식과 간염표면항원에 대한 단일클론항체 생산에 미치는 영향)

  • 전복환;조의철김동일
    • KSBB Journal
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    • v.5 no.4
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    • pp.365-371
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    • 1990
  • To improve the growth of mouse hybridoma 2c3.1 secreting anti-Hepatitis B surface antigen monoclonal antibody (anti-HBsAg MAb), we had constructed an enriched medium and observed the effects of fetal bovine serum and serum-free supplements including human serum albumin, 'insulin and transferrin', and monoethanolamine. For further enhancement of growth, the concentrations of two major energy sources, glucose and glutamine, were strengthened with various ratios in the enriched medium. Maximum cell growth and monoclonal antibody production obtained in various ratios of glucose/glutamine with an inoculation concentration of 2$\times$105 cells/ml were 0.73$\times$106-4.62$\times$106 cells/ml and 65.1-422.6 $\mu\textrm{g}$/ml, respectively. Glutamine was round to be a major energy source and a limiting nutrient in comparison to glucose for 2c3.1 cell cultivation in enriched media with low serum.

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Development of Transient Gene Expression System using Seedlings

  • Choi, Jang-Won;Park, Hee-Sung
    • Journal of agriculture & life science
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    • v.45 no.6
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    • pp.193-199
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    • 2011
  • An efficient transient expression system has been developed and characterized for the production of foreign genes in seedlings. The seedlings can be easily produced from commercial seeds used for vegetable sprouts. In principal, a chemical abrasive was employed to generate wounds in seedlings prior to vacuum-infiltration with Agrobacterium tumefaciens bearing the target gene. This optimized chemical wounding-assisted agro-infiltration process resulted in up to 15-fold increase in $\beta$-glucuronidase (GUS) enzyme activity. This procedure has been used efficiently to express hepatitis B surface antigen (HBsAg) protein in a transient mode. Therefore, seedlings with proper wounds can be suggested as a convenient tool for the production of useful recombinant proteins.

Production of Recombinant Humanized Anti-HBsAg Fab Fragment from Pichia pastoris by Fermentation

  • Deng, Ning;Xiang, Junjian;Zhang, Qing;Xiong, Sheng;Chen, Wenyin;Rao, Guirong;Wang, Xunzhang
    • BMB Reports
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    • v.38 no.3
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    • pp.294-299
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    • 2005
  • In this report, we describe the high-yield secretory expression of the recombinant human anti-HBsAg Fab fragment from Pichia pastoris that was achieved by co-integration of the genes encoding the heavy and light chains (both under the control of alcohol oxidase promoter) into the genome of the yeast cells. The fed-batch fermentations were carried out in a 5 L scale. Both chains of the Fab were successfully expressed upon methanol induction. The absorbance ($OD_{600}$) of the broth can reach 350~500 at the end of fed-batch phase. After the induction, the expression level of the recombinant Fab (soluble) reached 420~458 mg/L. The recombinant Fab fragment was purified from the crude culture supernatant by ion exchange chromatography and the purity of the recombinant Fab fragment was over 95%. The affinity activities of the crude fermentation supernatant and the purified Fab were analyzed by indirect ELISA, which showed that the purified recombinant Fab fragment had high affinity activity with hepatitis B surface antigen.

Seroepidemiologic Study of Hepatitis B Virus (HBV) and Hepatitis C Virus (HCV) Infection among Koreans in Rural Area, Korea (농촌지역의 B형, C형 간염에 관한 혈청역학적 연구)

  • Shin, Hai-Rim;Kim, Joon-Youn;Song, Jue-Bok;Suh, Byung-Seong;Kim, Jung-Man;Lee, Bu-Ok;Kim, Su-Yang;Ryang, Mu-Guan;Lee, Chae-Un;Chun, Jin-Ho;Yoo, Keun-Young;Ahn, Yoon-Ok
    • Journal of Preventive Medicine and Public Health
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    • v.30 no.1 s.56
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    • pp.17-29
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    • 1997
  • To estimate the prevalence of hepatitis B virus (HBV) and hepatitis C virus (HCV) infection and to determine associated risk factors, a population-based seroepidemiologic study was carried out. In 1993, a health examination survey of the population was carried out in rural area known to have a high incidence of liver cancer. The study population were those who volunteered to participate in a health survey over 10 years of age. Examinees were interviewed by specially trained staffs. Sera from 1,033 study subjects were tested for hepatitis B surface antigen (HBsAg) by .everse passive hemagglutinin (RPHA) estimation and for hepatitis C virus antibody (anti-HCV) by 2nd generation passive hemagglutinin (PHA) estimation. The age and sex standardized prevalence of HBsAg was 6.3% which was similar to national average, but that of anti-HCV was 5.1% which was 4 to 5 times higher than that of blood donors or other health examinees in Korea. In a multivariate analysis, transfusion history, surgical operative history, and acupuncture history were not associated with HBsAg positivity. In contrast, acupuncture history (adjusted odds ratio[OR]=2.2 : 95% Confidence interval[CI] 1.0-4.7) and surgical operative history(adjusted OR=2.0 : 95% CI 1.0-4.1) were associated with anti-HCV positivity. The present study suggest that there is an highly endemic area of HCV infection in Korea and probably this endemicity is associated with a parenteral source of HCV infection other than blood transfusion.

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Knowledge and Practice of College Entrants Toward Hepatitis B (일부 대학신입생들의 B형 간염에 대한 지식과 실천)

  • Chi, Sung-Ai;Chang, Kyou-Chull;Park, Jong-Yoon
    • Journal of Preventive Medicine and Public Health
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    • v.21 no.1 s.23
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    • pp.31-46
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    • 1988
  • In order to designate a present status necessary for establishment of preventive measures and guidelines of health education against hepatitis B in the course of secondary school education, knowledge and practice toward hepatitis B virus infection was surveyed by a questionnare method on total of 4,855 college entrants in the academic year of 1987 and analyzed the data collected using IBM PC(Trigem 88-II) with SAS package program. About two per cent of college entrants had past history of HBV infections not showing any difference between both sexes and geographical regions. About one third(33.7%) of total students had tested hepatitis B surface antigen(HBsAg), only 4% had tested hepatitis B surface antibody(HBsAb) and vaccination rate amounted to 24.6%, one fourth of total subjects. Both serological tests and vaccination were most commonly performed during adolescence, showing higher rates in female students than in male students. The rates also seemed to be higher in those from urban cities than those from rural cities. Students who had acquired correct knowledge that hepatitis B was infected by virus were amounted to 78.5% of college entrants, and remaining 21.5% had misunderstood that rickettsia, bacteria, fungi or parasites were causal agents. Female students were better aware of the causal agents than male students but there was no difference between places of growth. As for mode of transmission of HBV, 51.5% of male students and 47.7% of female students had correct knowledge. A very few student had known that fact that HBV was transmitted by body fluids such as tear(6.9%), nasal discharge(10.1%) and semen or vaginal secretion(19.2%) and majority(75%) of students had misunderstood that hepatitis B virus would be transmitted per os through food ingestion. Approximately one half(48.9%) of college entrants had knew correctly whom to be vaccinated. Approximately one half of the students knew that hepatr;ma(57.8%) and liver cirrhosis(57.4%) might complicate with hepatitis B virus infection, whereas 12.0% of the students responded that bronchitis was one of the complications of hepatitis B infection. In summary of the above results, we highly recommend that health education program for eradication of hepatitis B virus infection should be introduced in curricula of secondary school education in this country.

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Hepatitis B, C, and D Virus Infections and AFP Tumor Marker Prevalence Among the Elderly Population in Mongolia: A Nationwide Survey

  • Dambadarjaa, Davaalkham;Mukhtar, Yerkyebulan;Tsogzolbaatar, Enkh-Oyun;Khuyag, Ser-Od;Dayan, Angarmurun;Oyunbileg, Nandin-Erdene;Shagdarsuren, Oyu-Erdene;Nyam, Gunchmaa;Nakamura, Yosikazu;Takahashi, Masaharu;Okamoto, Hiroaki
    • Journal of Preventive Medicine and Public Health
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    • v.55 no.3
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    • pp.263-272
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    • 2022
  • Objectives: Infections with hepatitis B, C, and D virus (HBV, HCV, and HDV) are a major public health problem and lead to serious complications such as cirrhosis and hepatocellular carcinoma. We aimed to determine the seroprevalence of hepatitis B surface antigen (HBsAg), anti-HCV, anti-HDV immunoglobulin G, alpha-fetoprotein (AFP), and dual and triple hepatitis virus infections in Mongolia. Methods: A total of 2313 participants from urban and rural regions were randomly recruited for this cross-sectional study. A questionnaire was used to identify the risk factors for hepatitis virus infections, and the seromarkers were measured using immunoassay kits. Results: Among all participants, the prevalence of HBV, HCV, and HDV was 15.6%, 36.6%, and 14.3%, respectively. The infection rates were significantly higher in females and participants with a lower education level, rural residence, older age, and a history of blood transfusion. HBV and HCV co-infection was found in 120 (5.2%) participants and HBV, HCV, and HDV triple infection was detected in 67 (2.9%) participants. The prevalence of elevated AFP was 2.7%, 5.5%, and 2.6% higher in participants who were seropositive for HBsAg (p=0.01), anti-HCV (p<0.001), and anti-HDV (p=0.022), respectively. Elevated AFP was more prevalent in participants co-infected with HBV and HCV (5.8%, p=0.023), HBV and HDV (6.0%, p<0.001), and triple-infected with HBV, HCV, and HDV (7.5%) than in uninfected individuals. Conclusions: Nearly half (49.8%) of the study population aged ≥40 years were infected with HBV, HCV, or HDV, and 22.4% had dual or triple infections.

The Comparison of Results Among Hepatitis B Test Reagents Using National Standard Substance (국가 표준물질을 이용한 B형 간염 검사 시약 간의 결과 비교)

  • Lee, Young-Ji;Sim, Seong-Jae;Back, Song-Ran;Seo, Mee-Hye;Yoo, Seon-Hee;Cho, Shee-Man
    • The Korean Journal of Nuclear Medicine Technology
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    • v.14 no.2
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    • pp.203-207
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    • 2010
  • Purpose: Hepatitis B is infection caused by Hepatitis B virus (HBV). Currently, there are several methods, Kits and equipments for conducting Hepatitis B test. Due to ununiformed methods, it would cause some differences. To manage these differences, it needs process evaluating function of test system and reagent using particular standard substance. The aim of this study is to investigate tendency of RIA method's reagent used in Asan Medical Center through comparing several other test reagents using national standard substance. Materials and Methods: The standard substance in National Institute of Food and Drug Safety Evaluation's biology medicine consists of 5 things, 4 antigens and 1 antibody. We tested reagents using A, B company's Kits according to each test method. All tests are measured repeatedly to obtain accurate results. Results: Test result of "HBs Ag Mixed titer Performance panel" is obtained match rate compared S/CO unit standard with RIA method and EIA 3 reagents, CIA 2 reagents is that company A's reagent is 94.4% (17/18), 83.3% (15/18), B is 88.9% (16/18), 77.8% (14/18). Test result of "HBs Ag Low titer Performance panel" is obtain that EIA 2 reagents is shown 7 posive results, CIA 3 reagents is 11, and RIA method's company A's reagent is 3, B is 2 of 13 in low panel. "HBV surface antigen 86.76 IU/vial" tested dilution. A is obtain positive results to 600 times(0.14 IU/mL), B is 300 times (0.29 IU/mL). Case of "HBV human immunoglobulin 95.45 IU/vial", A is shown positive result to 10,000 times (9.5 mIU/mL) and B is 4,000 times (24 mIU/mL). Test result of "HBs Ag Working Standards 0.02~11.52 IU/mL" is shown that Company A's kit concentration level was 0.38IU/mL, company B was 2.23 IU/mL and higher level of concentration was positive results. Conclusion: When comparing various test reagents and RIA method according to National Standard substances for Hepatitis B test, we recognized that there were no significant trends between reagents. For hepatitis B virus antigen-antibody titers even in parts of the test up to 600 times the antigen, antibodies to 10,000 times the maximum positive results could be obtained. Therefore, we confirmed that results from Asan Medical Center are performed smoothly by reagents and system for hepatitis B virus test.

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Quantitative Analysis of Transient Expression in Tah Tasai Chinese Cabbage (Brassica campestris var. narinosa) Seedlings Following Agrobacterium-Mediated Transformation (다채 (Brassica campestris var. narinosa) 유묘의 형질전환 및 일시발현의 정량적 분석)

  • Shin Dong-Il;Park Hee-Sung
    • Journal of Plant Biotechnology
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    • v.32 no.4
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    • pp.275-279
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    • 2005
  • Tah tasai chinese cabbage (Brassica campestris var. narinosa), a vegetable plant popularly consumed as several-days-old seedlings in oriental countries, can be easily cultivated using a simple appliance. We demonstrated that Agrobacterium-mediated transformation via vacuum infiltration (agroinfiltration) resulted in a successful transient GUS gene expression in tah tasai chinese cabbage seedlings. Pre-germinated seeds were found to be more susceptible to Agrobacterium infection than one-day-old or two-days-old seedlings. We also demonstrated that hydrogen peroxide (HPO) treatment increased GUS expression especially for two-days-old seedlings. In ELISA using seedlings transformed with hepatitis B surface antigen (HBsAg) DNA by agroinfiltration, HBsAg protein synthesis increased more than two folds by HPO treatment to two-days-old seedlings in comparison to the mock-treated pre-germinated seeds.

Agrobacterium-Mediated Transformation of Flammulina velutipes with NaOH Treatment (NaOH처리에 의한 Agrobacterium이용 팽이균사체 형질전환)

  • Shin, Dong-Il;Park, Hee-Sung
    • The Korean Journal of Mycology
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    • v.39 no.3
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    • pp.235-238
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    • 2011
  • Agrobacterium harboring vector pCHBs with hygromycin phosphotransferase(hph) and hepatitis B virus surface antigen(HBsAg)gene was transformed into the mycelial culture of Flammulina velutipes. In particular, mild NaOH solution was treated to the mycelia before Agrobacterium infection step. This was purposed to generate putative surface wounds in the mycelial cell walls. The results showed that hygromycin-resistant($hyg^r$) mycelia could be obtained only from NaOH-treated mycelia but not from intact mycelia. The integration of $hyg^r$ gene in fungal genome was confirmed by PCR. In addition, a single transgene integration and heterologous protein expression in F. velutipes could be verified by Southern blot hybridization and western blot analysis, respectively. This study demonstrated an efficient tool for the Agrobacterium-mediated transformation of F. velutipes mycelia.

Seroprevalence of antibody to the hepatitis C virus in methamphetamine abusers (Methamphetamin 남용입원환자들의 C형간염 항체 양성률에 관한 조사)

  • Kim, Jin-Kyoo;Lee, Ji-Ho;Cho, Byung-Mann;Lee, Soo-Il
    • Journal of Preventive Medicine and Public Health
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    • v.24 no.4 s.36
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    • pp.465-472
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    • 1991
  • This study was designed to estimate the prevalence of hepatitis C virus(HCV) infection in drug abusers. The subjects were 141 inpatients who had been admitted to a general hospital with the symptoms and signs of methamphetamine intoxication. Seroprevalence of antibody to the hepatitis C virus(anti-HCV) was 60.3%(85/141) and it was higher in the group with increased frequency and duration of drug abuse, but such a relationship was not found in the seroprevalence of hepatitis B surface antigen(HBsAg). These findings suggested the possibility of high prevalence of HCV infection in methamphetamine abusers, and the importance of repetitive percutaneous injection in the transmission of HCV infection.

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