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

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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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Enhanced mucosal and systemic immune responses by mucosally administered hepatitis B surface antigen: effects of vaccine delivery vehicles and adjuvants

  • Park, Jeong-Sook;Kim, Chong-Kook
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.121-121
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    • 2003
  • The purpose of this study is to investigate the effect of mucosal vaccine delivery vehicles and adjuvants on the local and systemic antibody responses following mucosal immunization of mice with hepatitis B surface antigen (HBsAg). Mice were immunized on days 0 and 21 by administration of hepatitis B surface antigen B (HBsAg) into the vagina. HBsAg was delivered in saline or poloxamer(Pol)-based vehicle containing mucoadhesive polycarbophil (PC). (omitted)

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Physicochemical Properties of Recominant 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.521-526
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    • 1998
  • The physicochmical properties of recombinant hepatitis B surface antigen (r-HBsAg), which was expressed in C127 mammalian cell were studied. Using roller bottle culture in DMEM supplemented with fetal bovine serum, 10-15 mg/L of r-HBsAg was produced with about 31% of purification yield. The purity of r-HBsAg by HPLC was 99.8% and electron microscopic examination showed homogeneous spherical particle with 22 nm in diameter, a morphological characteristic of HBsAg. The density of r-HBsAg by CsCI density gradient method was 1.19g/ml and the isoelectric point by Mono $P^{TM}$ HR 5/20 column was 4.6. The analysis of subunit protein pattern using SDS-PAGE followed by scanning densitometry gave 81.3% of S protein and 18.7% of pre-S protein. fluorophore-assisted-carbohydrate-electrophoresis analysis showed the relative amount of carbohydrate to protein was 1.7% and it smajr component was N-acetyl glucosamine, which was about 39% of total carbohydrate. The relative amount of lipid to protein determined by vanillin phosphoric acid method was 32.5% and its major component was phospholipid, which was about 70% of total lipid. The physicochemical properties of C127 mammalian cell-derved r-HBsAg are similar to those of p-HBsAg, suggesting that the r-HBsAg can be used in developing a new preventive vaccine against hepatitis B.

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A Study on the Trend of Changes in the Positive Rate of Hepatitis B Surface Antigens and Antibodies over the Last 9 Years in One Health Care Center

  • Kang-Sil Lee;Sang-Ha Kim;Young-Bin Yu;Young-Kwon Kim
    • Biomedical Science Letters
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    • v.29 no.4
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    • pp.314-320
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    • 2023
  • The purpose of this study was to provide basic data on hepatitis B infection control in the community through the results of the hepatitis B surface antigen and antibody tests conducted at the Cheongyang-County Health Medical Center. From 2012 to 2020, we retrospectively analyzed the HBsAg, HBsAb, HBeAg, HBeAb, and HBV DNA results of 7,329 hepatitis B-related testers. Among 7,329 subjects, the HBsAg positivity rate was 1.7%, and the positivity rate according to age was the highest at 4.4% in their 30s, 4.2% in their 40s, 4.1% in their 50s, 2.0% in their 60s, 1.9% in their 70s and over, and 10 it was shown in the order of 0.3% from less than large. The HBsAb positivity rate was 43.1% for men, 38.2% for men, and 46.7% for women (P<0.001). To summarize the above results, for infection control of hepatitis B in Cheongyang-County, hepatitis surface antigen proton management is required for those in their 30s or older, and it is thought that efforts to acquire immunity are necessary for those in their 20s or younger.

A Case Report of HBsAg Seroclearance in Chronic Hepatitis B Patient

  • Lee, Hyeok Jae;Lee, Min-Hyeok
    • Korean Journal of Clinical Laboratory Science
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    • v.44 no.3
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    • pp.142-146
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    • 2012
  • Hepatitis B surface antigen (HBsAg) seroclearance is a rare event in chronic hepatitis B virus (HBV) infection which acquires the disease early in life. A case study have examined with asymptomatic chronic hepatitis B carrier who exhibits HBsAg seroclearance in anti-HBe positive. We comprehensively studied the biochemical, virological and clinical aspects of a patient with HBsAg seroclearance. Liver biochemistry, serological markers, serum HBV DNA levels, and development of clinical complications were monitored. Mutation of hepatitis B virus is suspected serum HBsAg detected by the HBsAg assay systems of VITROS (OrthoClinical Diagnostics, USA), AxSYM (Abbott Laboratories, USA), Elecsys (Roche Diagnostics, Germany) and ADVIA Centaur (Bayer Diagnostics, USA). These four immunoassays showed negative results. Also, the patient had undetectable serum HBV DNA. Therefore, no mutation within the "a" determinant of HBsAg, which might escape detection from HBsAg immunoassay were found. Natural seroclearance was confirmed.

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Sex Differences Associated With Hepatitis B Virus Surface Antigen Seropositivity Unwareness in Hepatitis B Virus Surface Antigen-positive Adults: 2007-2012 Korea National Health and Nutrition Examination Survey

  • Jang, Suk-Yong;Jang, Sung-In;Bae, Hong-Chul;Shin, Jaeyong;Park, Eun-Cheol
    • Journal of Preventive Medicine and Public Health
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    • v.48 no.2
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    • pp.74-83
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    • 2015
  • Objectives: To examine the sex-specific factors associated with being unaware of one's hepatitis B virus surface antigen (HBsAg) seropositivity status in a large, HBsAg-positive population of Koreans. Methods: In total, 1197 subjects aged 19 years or older who were HBsAg-positive according to data from the 2007-2012 Korea National Health and Nutrition Examination Survey were included. Subjects were considered unaware of their HBsAg seropositivity status if they answered that they had no knowledge of being previously infected by the hepatitis B virus (HBV) or diagnosed with HBV hepatitis. Multivariate Poisson regression models with robust variance estimate were used to assess the significance of the variables using weighted frequencies. Results: The majority (77.8%) of HbsAg-positive Korean adults (females, 81.9%; males, 74.6%) were unaware of their HBsAg seropositivity status. We found that sex (female: prevalence ratio [PR] 1.19), household income (low: PR, 1.15), marital status (never married: PR, 1.18), self-rated health (moderate: PR, 1.14; good: PR, 1.12), and alcohol use (at least 2-3 times/wk: PR, 1.21) were associated with being unaware. In females, age (50 to 59 years: PR, 1.29; ${\geq}70$ years: PR, 1.30), household income (low: PR, 1.37; middle-low: PR, 1.24), and marital status (never married: PR, 1.33) were associated with being unaware. In males, self-rated health (moderate: PR, 1.14; good: PR, 1.21) and alcohol use (at least 2-3 times/wk: PR, 1.21) were associated with being unaware. Conclusions: Factors related to the socioeconomic status of females and the health-related behaviors of males were found to be associated with being unaware of one's HBsAg seropositivity status.

Conformation of Group "a" Epitope in Hepatitis B Surface Antigen

  • Chun, Mun-Ho;Park, Won-Bong;Bok, Jin-Woo;Kim, Ha-Won;Choi, Eung-Chil;Kim, Byong-Kak
    • Archives of Pharmacal Research
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    • v.15 no.4
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    • pp.347-355
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    • 1992
  • To elucidate structure of group "a" epitope, mouse antibodies that express idiotype monoclonal antibody and anti-idiotype monoclonal antibody against the group specific "a" determinant were purified by hydroxyapatite column. To obtain hepatitis B surface antigens (HBsAg). HBsAg positive blood was sequencially purified by ammonium sulfate precipitation, hydroxyapatite, sepharose 4B column chromatography and ultracentrifugation. The major protein (p25) and glycoprotein (gp30) of HBsAg were isolated by concanavalin-A-sepharose 4B. The ability of p25-gp30 among the HBsAg to inhibit the idiotype-anti-idiotype reaction was dependent on conformation, since reduced and alkylated p25-gp30 virtualy lost their inhibitory capacity when compared to native HBsAg. The data suggest that hepatitis B antigen is a conformational antigen critically dependent upon the disulfide bonds of p25-gp30.

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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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Antibody productivity of HBsAg containing both preS2 and S regions expressed in Chinese hamster ovary cells (Chinese hamster ovary세포에서 발현된 pres2 및 S부위 함유 HBsAg의 항체유발능)

  • 정성균;박정민;이상봉;박동우;김동연;김기호;김홍진
    • YAKHAK HOEJI
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    • v.45 no.6
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    • pp.708-714
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    • 2001
  • Many studies have provided evidences that hepatitis B surface antigen (HBsAg) including preS region could be an ideal candidate for a new hepatitis B virus (HBV) vaccine with higher efficacy. We established CHO cell lines, IY-CHO-2 and IY-CHO-11 expressing high levels of HBsAg containing preS2 and S protein by stable transfection method. These cell lines expressed the correct size (about 1 kb in length) of HBsAg mRNA as expected. The purified protein from the culture supernatants of the clones showed the same sizes as those expressed in native hepatitis B virus (24 kDa, 27 kDa, 34 kDa and 36 kDa). Antibody productivity of CHO-derived HBsAg protein at lower dose challenge was higher than the protein containing S region alone expressed in yeast system. These results indicate that CHO-derived HBsAg protein containing preS2 and S region can be effectively used for a better immune response as a HBV vaccine.

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Quantitative Assay of Hepatitis B Surface Antigen by Using Surface Plasmon Resonance Biosensor

  • Hwang, Sang-Yoon;Yoo, Chang-Hoon;Jeon, Jun-Yeoung;Choi, Sung-Chul;Lee, Eun-Kyu
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.4
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    • pp.309-314
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    • 2005
  • We performed a basic experiment for the rapid, on-line, real-time measurement of hepatitis B surface antigen using a surface plasmon resonance biosensor. We immobilized anti­HBsAg (hepatitis B surface antigen) polyclonal antibody, as a ligand, to the dextran layer on a CM5 chip surface that had previously been activated by N-hydroxysuccinimide. A sample solution containing HBsAg was fed through a microfluidic channel, and the reflecting angle change due to the mass increase from the binding was detected. The binding characteristics between HBsAg and its polyclonal antibody followed the typical monolayer adsorption isotherm. When the entire immobilized antibody had interacted, no additional, non-specific binding occurred, suggesting the immunoreaction was very specific. The bound antigen per unit mass of the antibody was independent of the immobilized ligand density. No significant steric hindrance was observed at an immobilization density of approximately $17.6 ng/mm^2$. The relationship between the HBsAg concentration in the sample solution and the antigen bound to the ligand was linear up to ca. $40{\mu}g$/mL. This linearity was much higher than that of the ELISA method. It appeared the anti­gen-antibody binding increased as the immobilized ligand density increased. In summary, this study showed the potential of this SPR biosensor-based method as a rapid, simple and multi­sample on-line assay. Once properly validated, it may serve as a more efficient method for HBsAg quantification for replacing the ELISA.