• 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.

• 대한핵의학회:학술대회논문집
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• 1999.11a
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• pp.26.2-26.2
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• 1999

• 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.

• Journal of Preventive Medicine and Public Health
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• v.21 no.2 s.24
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• pp.329-339
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• 1988

The study is carried out to investigate the effect of drinking and smoking for the activities of aspartate aminotransferase(AST, or GOT) and alanine aminotransferase(ALT or GPT), from December 25, 1986 to April 30, 1987. The male physical examinees for employment, 900 who had visited to the Taegu Medical Center were subjected. And the positive cases of HBs-Ag, Anti-HBs and skin test for Clonorchis sinensis were excluded. The general characters of drinking and smoking pattern were introduced by interview with questionnaire provided for. In drinking cases, the longer duration was significantly effected the higher rate of abnormality in AST and ALT level. But the amount and the frequency were not. It was not appeared effects by mackgulri which is a Korean traditional wine and small amount of beers. In smoking cases, also same pattern. The age was related in all cases. By the way, when the effect is related the positive results with other factors: HBs-Ag, Anti-HBs, skin test for Clonorchiasis and harmful occupational history, it is higher abnormal rate of AST and ALT in the duplicated cases with two factors or more. Particularly in HBs-Ag positive cases, those who had smoking was the highest in rate of abnormality, and drinking was the follows. In correlation matrix among seven factors; HBs-Ag, age, drinking amount, drinking period, drinking frequency, smoking amount and smoking period, correlation coefficient was significant between the abnormal rate and to with age, drinking period, smoking period, and smoking amount.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.5
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• pp.362-365
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• 2000

A rapid immunochromatographic assay kit using whole blood to screen hepatitis B surface antigen was developed and evaluated by using sera from 240 patients. The reference diagnosis was based on the results obtained with GENEDIA Anti-HBs Rapid kit which is very similar to the above kit except for the use of serum. The test demonstrated a good correlation with the reference immunochromatographic assay kit, that is, the sensitivity and the specificity of the kit was 100%, respectively. The rapid test kit using whole blood should be more convenient and useful for the diagnosis of hepatitis B virus because the kit does not need machines and time to prepare serum. In addition, this kit is safe from inadvertent infection during sample treatment because the blood is sterilized with hydrogen peroxide, eliminates the procedure required to prepare serum and reduces the possibility of exposure to infectious agents.

• Journal of Preventive Medicine and Public Health
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• v.27 no.4 s.48
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• pp.747-761
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• 1994

Workers' periodic health examination is the main tools used to manage the health problems of most workers in Korea. The most common health problem found in workers' periodic health examination is liver disorder. Liver disorder is also one of the most common health problems in general population and one of the leading causes of mortality in adult population. Regulation proposed by government (No. 207, Ministry of Labor, 1992) defines the criteria for selection of workers with the liver dysfunction for further evaluative examination and the examination items used for diagnosis of the workers with liver dysfunction. This study was designed to evaluate the proficiency of each examination items presently defined in Regulation and propose the optimal examination items for detection of the liver disorders found by workers' periodic health examination. Study subjects are 186 workers with abnormal liver function tests in screening examination of workers' periodic health examination. Questionnaire survey including past history of liver disorder, drinking history, height and weight was done. Physical examination by physician, routine test items defined by Regulation (SGOT, SGPT, $\gamma$-GTP, protein, albumin, total and direct bilirubin, alkaline phosphatase, $\alpha$-feto protein, HBsAg and anti-HBs), anti-HCV antibody test and liver ultrasonography were done. Results are as follows; 1. Result of evaluative examination utilizing only the items defined in Regulation was; There were 75 workers with suspected live. disorder(40.3%), 63 with no liver dysfunction (33.9%), 13 with suspected hepatitis B(7.0%), 10 workers with hepatitis B(5.4%), 10 workers with hepatitis B carrier state(5.4%), 10 with alcoholic liver disorders(5.4%), 5 with fatty liver(2.7%). When alternative diagnostic criteria applying additional examination items (drinking history, body mass index, anti-HCV antibody and ultrasonography) diagnosability of liver disorder was increased. When all four items were included, final results were; 23 workers (17.8%) with hepatitis B (10 carriers, 13 suspects and 10 hepatitis B), 10 (5.4%) with hepatitis C(4 carriers, 5 suspects and 1 hepatitis C), 13(7.0%) with alcoholic liver disorder, 45(24.2%) with fatty liver (40 suspects, 5 fatty liver), 410%) with suspected liver disorders and 44 (23.7%) with normal liver. 2. Of examination items defined by Regulation, only SGOT, SGPT, $\gamma$-GTP and HBsAg were significantly different in abnormal rate and mean value, and all other laboratory findings did not showed significant difference between two groups. Drinking history, body mass index and anti-HCV antibody test which are the items that authors included in this study, also showed significant difference between two groups. Utilization of body mass index (BMI) for abnormal liver function group in diagnosis of fatty liver had high specificity (97.6%) but sensitivity (22.3%) was low. Therefore we suggest that SGOT, SGPT, $\gamma$-GTP, HBsAg, alcohol drinking history, BMI and anti-HCV Ab were useful for diagnosis of liver disorders among worker's periodic health examination.

• Pediatric Infection and Vaccine
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• v.11 no.1
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• pp.73-80
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• 2004

Purpose : The serial clinical findings, biochemical results, and serological hepatitis B virus(HBV) markers in Korean children with chronic HBV infection were analyzed to determine the relationships among these factors. Methods : Ninety children have been chosen from those who have visited to the Department of Pediatrics at St. Vincent's Hospital in The Catholic University of Korea from July 1st, 1995 to June 30th, 2000. The sample patients were followed up for over six months. HBV markers and liver function tests were all performed. Results : All children were asymptomatic at presentation. Eighty-three percent of the children had a history of chronic HBV infection in their families. Eighty-one percent were HBeAg positive, 16% were anti-HBe positive, while 3% were all HBeAg and anti-HBe negative. The prevalence of HBeAg among three age groups : 0~5; 6~10; and 11~15 year-old was 90%, 96% and 61% respectively. The prevalence of HBeAg in less than 10 year-old group was significantly higher than 11~15 year-old group(P=0.001). Serum ALT levels were within 40 IU/L in 64% children, 41~80 IU/L in 17%, 81~200 IU/L in 10%, and beyond 201 IU/L in 9%. The percentage of abnormality of ALT levels in HBeAg positive patients was significantly higher than that of HBeAg negative(P=0.036). Eleven of the 73 HBeAg positive children lost their HBeAg and seroconverted to anti-HBe. In these cases, all had transient elevations in ALT levels before HBeAg seroconversions. The annual rates of spontaneous seroconversion of HBeAg and HBsAg were 9.7% and 0.6%, respectively. Conclusion : Recognition of the dynamics of these changes in viral markers and biochemical findings is needed in the selection and evaluation of therapeutic regimens, establishment of treatment, and calling for controlled trials with adequate follow-up. The hepatitis B carrier state may be asymptomatic in children however, continued surveillance of carriers is important to determine the individual adverse prognostic factors of chronic HBV infections.

• Journal of Yeungnam Medical Science
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• v.13 no.2
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• pp.272-278
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• 1996

Four subtypes of hepatitis B surface antigen are useful in the epidemiologic studies of the route of virus transmission and clinical significance of simultaneous occurance of hepatitis B surface antigen and antibody to hepatitis B surface antigen in the same serum as well as useful marker for population migration. The sera were obtained from 214 HBs Ag positive patients who are diagnosed as chronic liver disease and following up in the Yeungnam university hospital. The subtypes were determined by solid-phase sandwich EIA using monoclonal antibodies. Among 214 specimens, the subtype adr was 93.9%, adw was 2.8%, ayr was 0.9%, ar was 0.9%, adwr was 1.4% and ayw was not detected. There were no correlation between subtype pattern and disease. In summary, the subtype adr was prominent in our study and the difference of subtype pattern by severity of disease was not significant. However, to determine the prognostic value of HBs Ag subtype and relationship between subtype and disease progression, long-term follow up will be needed.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.6 no.2
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• pp.140-151
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• 2003

Purpose: We tried to evaluate the long term efficacy and positive predictive factors of interferon-alpha treatment in children with chronic hepatitis B. Methods: The study population included 113 children who received interferon therapy between May 1982 and July 2002 (20 years) for chronic hepatitis B in Department of Pediatrics, Yonsei University College of Medicine. Male to female ratio was 2.3 : 1 and the mean age at diagnosis was $11.1{\pm}4.1$ years old. Response to treatment was defined as normalization of alanine aminotransferase (ALT), disappearance of HBeAg and HBV-DNA Eighty two children responded while 32 did not. Interferon-alpha was given intramuscularly for 6 months at a dosage of $3{\times}10^6$ unit, 3 times weekly. In relapsed cases, lamivudine or interferon retreatment was done. Results: Seroconversion rate was 77.0% in terms of HBeAg, 74.3% in terms of HBV-DNA, and 80.5% in terms of ALT normalization after treatment. Seroconversion rate of both HBeAg and HBV-DNA was 72.6%. Analyzed by life table method, the effect of the treatment had been maintained over 10 years after cessation of therapy. Pre-treatment ALT level was the only significant positive predictive factor of response. Eleven cases (13.4%) relapsed, and 2 out of 3 showed response when treated with lamivudine and 1 out of 3 with interferon retreatment. Conclusion: Interferon-alpha showed significant efficacy in the treatment of chronic hepatitis B in our study. Further studies about the effect of interferon therapy on complications of hepatitis such as hepatocarcinoma, cirrhosis are warranted.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.23 no.4
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• pp.311-318
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• 2020

Chronic hepatitis B viral (HBV) infection remains a major health threat, especially in high-prevalence areas. Most infants infected by mother-to-infant HBV transmission become chronic carriers. In Taiwan, many important preventive interventions have been implemented to block the perinatal transmission of HBV in the past 35 years. The first nationwide universal HBV vaccination program was launched in Taiwan in July 1984. The three-dose HBV vaccine completion rate reached 98.1% in 2018. The prevalence of Hepatitis B surface antigen (HBsAg) decreased from 9.8% in pre-vaccinated period in 1984 to 0.5% in the vaccinated cohort in 2014. The incidence of hepatocellular carcinoma in children aged 6-9 years significantly declined from 0.52 to 0.13 per 100,000 children born before and after 1984, respectively. Furthermore, we have performed a maternal HBV screening program during pregnancy since 1984, with the screening rate peaked at 93% in 2012. The HBsAg- and HBeAg-seropositive rate in pregnant women declined from 13.4% and 6.4% in 1984-1985 to 5.9% and 1.0% in 2016, respectively. To closely control perinatal HBV infection, we have administered hepatitis B immunoglobulin immediately after birth and checked the serum level of HBsAg and anti-HBs in high-risk babies born to HBsAg-seropositive mothers, irrespective of their HBeAg status, since July 2019. We have also adopted short-term antiviral treatments such as tenofovir 300 mg daily in the third trimester for highly viremic mothers and reduced the perinatal infection rates from 10.7 to 1.5%. Through all these efforts, we expect to meet the global goal of eliminating HBV infection by 2030.