• 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.

• 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.

• KSBB Journal
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• v.15 no.2
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• pp.214-218
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• 2000

A hepatitis B Surface Antigen(HBsAg)-screening kit using immunochromatographic assay(ICA) method was developed by e employing two kinds of antibodies. One is mouse monoclonal anti-HBs for tracer antibody and the other is goat p이yclonal a anti-HBs for capture antibody. This capture antibody was immobilized on the surface of nitroceliulose(NC) membrane and the t tracer antibody was conjugated with g미d particles. When serum sample was added to the sample well, the $\infty$njugates d deposited in a dry state on the surface of glass fiber filter were reconstituted and then combined with HBsAg in serum. In 5 5 min after adding, the assay result was visible through the window, that is, the complexes composed of HBsAg and the c conjugates appeared as maroon line on the lower part of the NC membrane. The detection limit of the ICA kit was 2 ng/ml w when being tested with the reference HBsAg.

• 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.

• KSBB Journal
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• v.17 no.1
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• pp.20-25
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• 2002

We performed a basic experiment for rapid, on-line, real-time measurement of HBsAg by using a surface plasmon resonance biosensor to quantify the recognition and interaction of biomolecules. We immobilized the anti-HBsAg polyclonal antibody to the dextran layer on a CM5 chip surface which was pre-activated by N-hydroxysuccinimide for amine coupling. The binding of the HBsAg to the immobilized antibody was measured by the mass increase detected by the change in the SPR signal. The binding characteristics between HBsAg and its antibody followed typical monolayer adsorption isotherm. When the entire immobilized antibody was interacted, there was no additional, non-specific binding observed, which suggested the biointeraction was very specific as expected and independent of the ligand density. No significant steric hindrance was observed at 17.6 nm/$mm^2$ immobilization density. The relationship between the HBsAg concentration in the sample solution and the antigen bound to the chip surface was linear up to ca. $40\mu\textrm{g}$/mL, which is much wider than that of the ELISA method. It appeared the antigen-antibody binding was increased as the immobilized ligand density increased, but verification is warranted. This study showed the potential of this biosensor-based method as a rapid, simple, multi-sample, on-line assay. Once properly validated, it can serve as a more powerful method for HBsAg quantification replacing the current ELISA method.

• 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.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.231-234
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• 2001

We performed a basic experiment for rapid. on-line, real-time measurement of HBsAg by using a BIACORE biosensor, a chip-based sensor utilizing surface plasmon resonance technology to quantify the recognition and interaction of biomolecules. We immobilized an a -HBsAg antibody on a CM5 chip surface which was activated by N-hydroxysuccinimide for amine coupling with HBsAg, and measured the mass increase from the coupling. This study showed the potential of this biosensor-based method as a rapid, multi-sample, on-line assay. Once properly validated, it can serve as a more powerful method for HBsAg quantification.

• Annals of Laboratory Medicine
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• v.38 no.6
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• pp.578-584
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• 2018

Background: Accurate, rapid, and cost-effective screening tests for hepatitis B virus (HBV) and hepatitis C virus (HCV) infection may be useful in laboratories that cannot afford automated chemiluminescent immunoassays (CLIAs). We evaluated the diagnostic performance of a novel rapid automated fluorescent lateral flow immunoassay (LFIA). Methods: A fluorescent LFIA using a small bench-top fluorescence reader, Automated Fluorescent Immunoassay System (AFIAS; Boditech Med Inc., Chuncheon, Korea), was developed for qualitative detection of hepatitis B surface antigen (HBsAg), antibody to HBsAg (anti-HBs), and antibody to HCV (anti-HCV) within 20 minutes. We compared the diagnostic performance of AFIAS with that of automated CLIAs-Elecsys (Roche Diagnostics GmbH, Penzberg, Germany) and ARCHITECT (Abbott Laboratories, Abbott Park, IL, USA)-using 20 seroconversion panels and 3,500 clinical serum samples. Results: Evaluation with the seroconversion panels demonstrated that AFIAS had adequate sensitivity for HBsAg and anti-HCV detection. From the clinical samples, AFIAS sensitivity and specificity were 99.8% and 99.3% for the HBsAg test, 100.0% and 100.0% for the anti-HBs test, and 98.8% and 99.1% for the anti-HCV test, respectively. Its agreement rates with the Elecsys HBsAg, anti-HBs, and anti-HCV detection assays were 99.4%, 100.0%, and 99.0%, respectively. AFIAS detected all samples with HBsAg genotypes A-F and H and anti-HCV genotypes 1, 1a, 1b, 2a, 2b, 4, and 6. Cross-reactivity with other infections was not observed. Conclusions: The AFIAS HBsAg, anti-HBs, and anti-HCV tests demonstrated diagnostic performance equivalent to current automated CLIAs. AFIAS could be used for a large-scale HBV or HCV screening in low-resource laboratories or low-to middle-income areas.

• Korean Journal of Clinical Laboratory Science
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• v.47 no.2
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• pp.78-82
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• 2015

The purpose of this study was to survey the anti-HBs positivity and titers of antibody in current students who received nationwide vaccination against Hepatitis B virus which targeted infants in 1995. The subjects were 262 students in Gyeongnam province from April 2014 to October 2014. The positive rate of anti-HBs was 55.3% (145 people) and the negative rate of anti-HBs was 44.7% (117 people). Positivity was shown to be higher in women than men. However, there was no statistically significant difference. Of the HBV-vaccinated subjects, 117 (44.7%) had anti-HBs titer <10, which is judged to be negative, 126 (47.8%) had anti-HBs titer 10-499.9 mIU/mL, which is judged to be positive, and 22 (7.3%) had anti-HBs titer more than 500 mIU/mL. The rate of anti-HBs with lower titer (10-99.9 mIU/mL) was 62% in the positive group. Classifying the antibody titer according to age, the rate of anti-HBs positivity in titer with less than 100 mIU/mL was indicated to be 78.3% in cases of 19-20 year old and 46.7% in 21-22 year old, 52.3% in 23-24 year old. A case of the lower titer with 10-99.9 mIU/mL, showed significant difference according to age. As a result of research, the antibody titers is decreased depending on the passage of time. Hence, the checking of anti-HBs titer is needed after Hepatitis B vaccination and many healthy adults will need periodic boosters of hepatitis B vaccine to maintain production of antibody to hepatitis B surface antigen.

• 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.