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A phage-displayed cyclic peptide that interacts tightly with the immunodominant region of hepatitis B surface antigen

Tan WS, Tan GH, Yusoff K, Seow HF

J. Clin. Virol., 2005 Sep;34(1):35-41.
PMID: 16087122

The surface antigen (HBsAg) of hepatitis B virus (HBV) is highly conformational and generally evokes protective humoral immune response in human. A disulfide constrained random heptapeptide library displayed on the coat protein III of filamentous bacteriophage M13 was employed to select specific ligands that interact with HBsAg subtype ad. Fusion phages carrying the amino acid sequence ETGAKPH and other related sequences were isolated. The binding site of peptide ETGAKPH was located on the immunodominant region of HBsAg. An equilibrium binding assay in solution showed that the phage binds tightly to HBsAg with a relative dissociation constant (KDrel) of 2.9+/-0.9 nM. The phage bearing this peptide has the potential to be used as a diagnostic reagent and two assays for detecting HBsAg in blood samples are described.

Matched MeSH terms: Hepatitis B Surface Antigens/blood; Hepatitis B Surface Antigens/isolation & purification; Hepatitis B Surface Antigens/metabolism*; Hepatitis B Surface Antigens/chemistry
Antigenicity and immunogenicity of the immunodominant region of hepatitis B surface antigen displayed on bacteriophage T7

Tan GH, Yusoff K, Seow HF, Tan WS

J. Med. Virol., 2005 Dec;77(4):475-80.
PMID: 16254965

The immunodominant region of hepatitis B virus (HBV) located in the viral small surface antigen (S-HBsAg) elicits virus-neutralizing and protective antibodies. In order to develop an easy and inexpensive method to produce this region without the need for extensive purification, amino acid residues 111-156 of S-HBsAg were fused to the C-terminal end of the 10B capsid protein of T7 phage. Western blotting and ELISA confirmed the expression of the recombinant protein on the surface of the phage particles. The recombinant phage exhibited the antigenic and immunogenic characteristics of HBsAg, illustrating its potential as an immunological reagent and vaccine.

Matched MeSH terms: Hepatitis B Surface Antigens/analysis; Hepatitis B Surface Antigens/genetics; Hepatitis B Surface Antigens/immunology*; Hepatitis B Surface Antigens/chemistry
Fulltext Preventing Mother-to-child Transmission of Hepatitis B Virus - A Success Story Which Can Be Enhanced

Tan SS, Chua A

Med. J. Malaysia, 2013 Apr;68(2):103-4.
PMID: 23629552
Matched MeSH terms: Hepatitis B Surface Antigens*
Fulltext Enhanced cell disruption strategy in the release of recombinant hepatitis B surface antigen from Pichia pastoris using response surface methodology

Tam YJ, Allaudin ZN, Lila MA, Bahaman AR, Tan JS, Rezaei MA

BMC Biotechnol., 2012;12:70.
PMID: 23039947 DOI: 10.1186/1472-6750-12-70

Cell disruption strategies by high pressure homogenizer for the release of recombinant Hepatitis B surface antigen (HBsAg) from Pichia pastoris expression cells were optimized using response surface methodology (RSM) based on the central composite design (CCD). The factors studied include number of passes, biomass concentration and pulse pressure. Polynomial models were used to correlate the above mentioned factors to project the cell disruption capability and specific protein release of HBsAg from P. pastoris cells.

Matched MeSH terms: Hepatitis B Surface Antigens/genetics*; Hepatitis B Surface Antigens/isolation & purification; Hepatitis B Surface Antigens/metabolism
The full-length clone of cucumber green mottle mosaic virus and its application as an expression system for Hepatitis B surface antigen

Ooi A, Tan S, Mohamed R, Rahman NA, Othman RY

J. Biotechnol., 2006 Feb 24;121(4):471-81.
PMID: 16271415

A cucumber green mosaic mottle virus (CGMMV) full-length clone was developed for the expression of Hepatitis B surface antigen (HBsAg). The expression of the surface displayed HBsAg by the chimeric virus was confirmed through a double antibody sandwich ELISA. Assessment of the coat protein composition of the chimeric virus particles by SDS-PAGE analysis showed that 50% of the coat proteins were fused to the HBsAg. Biological activity of the expressed HBsAg was assessed through the stimulation of in vitro antibody production by cultured peripheral blood mononuclear cells (PBMC). PBMC that were cultured in the presence of the chimeric virus showed up to an approximately three-fold increase in the level of anti HBsAg immunoglobulin thus suggesting the possible use of this new chimeric virus as an effective Hepatitis B vaccine.

Matched MeSH terms: Hepatitis B Surface Antigens/biosynthesis*; Hepatitis B Surface Antigens/genetics; Hepatitis B Surface Antigens/immunology
Cloning, expression and display of the PreS domain of hepatitis B virus on filamentous bacteriophage M13

Kok WL, Yusoff K, Nathan S, Tan WS

J. Biochem. Mol. Biol. Biophys., 2002 Feb;6(1):55-8.
PMID: 12186783

The PreS domain of hepatitis B virus (HBV) is believed to be involved in virion assembly and attachment to a hepatocyte receptor during infection. In order to study the functions of this region, we fused it to the g3p protein of bacteriophage M13 that allows the fusion protein to be displayed at the tip of the filament. The fusion protein was detected by the anti-E tag antibody on a Western blot. The polypeptide in a soluble form was produced by transfecting a non-suppressor E. coli host cell with the recombinant phagemid. The soluble protein was detected in cytoplasm, in the periplasmic space and also in the medium. The functional display of the PreS domain would provide an alternative means to study its interactions with the nuleocapsid and hepatocytes.

Matched MeSH terms: Hepatitis B Surface Antigens/genetics*; Hepatitis B Surface Antigens/metabolism; Hepatitis B Surface Antigens/chemistry*
Trends in hepatitis B virus infection among blood donors in Kelantan, Malaysia: a retrospective study

Yousuf R, Rapiaah M, Ahmed SA, Rosline H, Salam A, Selamah S, et al.

Southeast Asian J. Trop. Med. Public Health, 2007 Nov;38(6):1070-4.
PMID: 18613548

The objective of this study was to determine the prevalence and trends in hepatitis B infection among blood donors attending the Transfusion Medicine Unit at the Hospital Universiti Sains Malaysia, Kelantan, Malaysia. A retrospective study was carried out by reviewing the results of HBsAg among blood donors for the years 2000 to 2004. During this period, 44,658 blood donors were studied. We noted that there was a significant difference in the prevalence of hepatitis B infection between regular and first time donors. There was also a decreasing trend noticed in both study groups. The mean prevalence was significantly different between first time (1.83%) and regular donors (0.45%) (p < 0.005). There is a need to improve public awareness programs to lower the incidence of hepatitis B infection in the general population and consequently first time blood donors. Future studies are also required to determine the trends and outcomes of these programs.

Matched MeSH terms: Hepatitis B Surface Antigens/blood; Hepatitis B Surface Antigens/isolation & purification
Prevalence of hepatitis B surface antigen and antibody among health care employees in Negri Sembilan, Malaysia, 1989

Tan TC, Vadivale M, Ong CN

Asia Pac J Public Health, 1992;6(3):134-9.
PMID: 1342800 DOI: 10.1177/101053959200600303

This study was based on a hepatitis B screening program conducted in one of the states in Malaysia in 1989. The majority (84.6%) of the 2986 health employees were screened. One quarter (25%) was found to have serological markers for the Hepatitis B Virus (HBV); 2.1% had Hepatitis B surface Antigen (HBsAg) and 22.8% had antibody to the Hepatitis B surface Antigen (anti-HBs). The occurrence of HBsAg was higher in ethnic Chinese (6.3%) compared to Malays (1.8%) and Indians (0.9%), even when analyzed by sex, but not with age, type of institution and geographical locality. The distribution of anti-HBs was higher with ethnic Chinese (41.6%), male sex (27.2%) and age. There was a wide variation of the prevalence of serological markers among occupations and increased relative risks of HBsAg were found among medical assistants (RR3.7; 95% CI 1.4-9.1) and laboratory staff (RR 3.2; 95% CI 1-8.8), and that of anti-HBs among medical assistants (RR 2.8; 95% CI 1.8-3.7). The variations of HBsAg among occupations by type of institutions was marginal while that of anti-HBs was higher among attendants and midwives in hospitals, medical assistants in health departments, and assistant nurses and dentists in dental centers. The patterns of distribution of serological markers of HBV among health staff reflect the situation in the community with high endemicity and resemble specific occupational factors noted in previous studies in the West.
Publication year is 1992-1993

Matched MeSH terms: Hepatitis B Surface Antigens/blood*; Hepatitis B Surface Antigens/immunology
Solution structure and in silico binding of a cyclic peptide with hepatitis B surface antigen

Muhamad A, Ho KL, Rahman MB, Uhrín D, Tan WS

Chem Biol Drug Des, 2013 Jun;81(6):784-94.
PMID: 23405984 DOI: 10.1111/cbdd.12120

A specific ligand targeting the immunodominant region of hepatitis B virus is desired in neutralizing the infectivity of the virus. In a previous study, a disulfide constrained cyclic peptide cyclo S(1) ,S(9) Cys-Glu-Thr-Gly-Ala-Lys-Pro-His-Cys (S(1) , S(9) -cyclo-CETGAKPHC) was isolated from a phage displayed cyclic peptide library using an affinity selection method against hepatitis B surface antigen. The cyclic peptide binds tightly to hepatitis B surface antigen with a relative dissociation constant (KD (rel) ) of 2.9 nm. The binding site of the peptide was located at the immunodominant region on hepatitis B surface antigen. Consequently, this study was aimed to elucidate the structure of the cyclic peptide and its interaction with hepatitis B surface antigen in silico. The solution structure of this cyclic peptide was solved using (1) H, (13) C, and (15) N NMR spectroscopy and molecular dynamics simulations with NMR-derived distance and torsion angle restraints. The cyclic peptide adopted two distinct conformations due to the isomerization of the Pro residue with one structured region in the ETGA sequence. Docking studies of the peptide ensemble with a model structure of hepatitis B surface antigen revealed that the cyclic peptide can potentially be developed as a therapeutic drug that inhibits the virus-host interactions.

Matched MeSH terms: Hepatitis B Surface Antigens/metabolism*; Hepatitis B Surface Antigens/chemistry
Display of hepatitis B virus PreS1 peptide on bacteriophage T7 and its potential in gene delivery into HepG2 cells

Tang KH, Yusoff K, Tan WS

J. Virol. Methods, 2009 Aug;159(2):194-9.
PMID: 19490973 DOI: 10.1016/j.jviromet.2009.03.015

Hepatitis B is a major public health problem worldwide which may lead to chronic liver diseases, cirrhosis and hepatocellular carcinoma. An interaction between hepatitis B virus (HBV) envelope protein, particularly the PreS1 region, and a specific cell surface receptor is believed to be the initial step of HBV infection through attachment to hepatocytes. In order to develop a gene delivery system, bacteriophage T7 was modified genetically to display polypeptides of the PreS1 region. A recombinant T7 phage displaying amino acids 60-108 of the PreS1 region (PreS1(60-108)) was demonstrated to be most effective in transfecting HepG2 cells in a dose- and time-dependant manner. The phage genome was recovered from the cell lysate and confirmed by PCR whereas the infectious form of the internalized phage was measured by a plaque-forming assay. The internalized phage exhibited the appearance of green fluorescent dots when examined by immunofluorescence microscopy. Surface modification, particularly by displaying the PreS1(60-108) enhanced phage uptake, resulting in more efficient in vitro gene transfer. The ability of the recombinant phage to transfect HepG2 cells demonstrates the potential of the phage display system as a gene therapy for liver cancer.

Matched MeSH terms: Hepatitis B Surface Antigens/genetics; Hepatitis B Surface Antigens/metabolism*
Genotyping of hepatitis B virus in Malaysia based on the nucleotide sequence of preS and S genes

Ong HT, Duraisamy G, Kee Peng N, Wen Siang T, Seow HF

Microbes Infect., 2005 Mar;7(3):494-500.
PMID: 15792534

Hepatitis B virus (HBV) has been classified into eight genotypes, designated A-H. These genotypes are known to have distinct geographic distributions. The clinical importance of genotype-related differences in the pathogenicity of HBV has been revealed recently. In Malaysia, the current distribution of HBV remains unclear. The aim of this study was to determine the genotypes and subtypes of HBV by using PCR, followed by DNA sequencing, as well as to analyse the mutations in the immunodominant region of preS and S proteins. The S gene sequence was determined from HBV DNA of four apparently healthy blood donors' sera and three sera from asymptomatic chronic hepatitis B carriers. Of this batch of sera, the preS gene sequence was obtained from HBV DNA from three out of the four blood donors and two out of the three chronic carriers. Due to insufficient sera, we had to resort to using sera from another blood donor to make up for the sixth DNA sequence of the preS gene. Based on the comparative analysis of the preS sequences with the reported sequences in the GenBank database, HBV DNA from two normal carriers was classified as genotype C. Genotype B was assigned to HBV from one blood donor and two hepatitis B chronic carriers, whereas HBV of one chronic carrier was of genotype D. Based on the S gene sequences, HBV from three blood donors was of genotype C, that of one blood donor and one chronic carrier was of genotype B, and the remaining, of genotype D. In the five cases where both preS and S gene sequences were determined, the genotypes assigned based on either the preS or S gene sequences were in concordance. The nature of the deduced amino acid (aa) sequences at positions 125, 127, 134, 143, 159, 161 and 168 of the S gene enabled the classification of these sequences into subtypes, namely, adrq+, adw2 and ayw2. The clustering of our DNA sequences into genotype groups corresponded to their respective subtype, that is, adw2 in genotype B, adrq in genotype C and ayw in genotype D. Analysis of the point mutations revealed that five of the sequences contained aa substitutions at immunodominant epitopes involved in B or/and T cell recognition. In conclusion, despite the low numbers of samples studied, due to budget constraints, these data are still worthwhile reporting, as it is important for the control of HBV infections. In addition, the genotype and mutational data obtained in this study may be useful for designing new treatment regimes for HBV patients.

Matched MeSH terms: Hepatitis B Surface Antigens/genetics*; Hepatitis B Surface Antigens/chemistry
Molecular and serological detection of occult hepatitis B virus among healthy hepatitis B surface antigen-negative blood donors in Malaysia

Hudu SA, Harmal NS, Saeed MI, Alshrari AS, Malik YA, Niazlin MT, et al.

Afr Health Sci, 2016 Sep;16(3):677-683.
PMID: 27917199

BACKGROUND: Occult hepatitis B infections are becoming a major global threat, but the available data on its prevalence in various parts of the world are often divergent.
OBJECTIVE: This study aimed to detect occult hepatitis B virus in hepatitis B surface antigen-negative serum using anti-HBc as a marker of previous infection.
PATIENT AND METHODS: A total of 1000 randomly selected hepatitis B surface antigen-negative sera from blood donors were tested for hepatitis B core antibody and hepatitis B surface antibody using an ELISA and nested polymerase chain reaction was done using primers specific to the surface gene (S-gene).
RESULTS: Of the 1000 samples 55 (5.5%) were found to be reactive, of which 87.3% (48/55) were positive for hepatitis B surface antibody, indicating immunity as a result of previous infection however, that does not exclude active infection with escaped mutant HBV. Nested PCR results showed the presence of hepatitis B viral DNA in all the 55 samples that were positive for core protein, which is in agreement with the hepatitis B surface antibody result.
CONCLUSION: This study reveals the 5.5% prevalence of occult hepatitis B among Malaysian blood donors as well as the reliability of using hepatitis B core antibody in screening for occult hepatitis B infection in low endemic, low socioeconomic settings.

Matched MeSH terms: Hepatitis B Surface Antigens/blood*; Hepatitis B Surface Antigens/genetics
Fulltext Hepatitis B surface antigen subtypes in hepatitis B seropositive subjects in University Hospital, Kuala Lumpur

Ng KP, Saw TL

Med. J. Malaysia, 1999 Sep;54(3):352-7.
PMID: 11045062

Hepatitis B surface antigen can be serologically defined as ayw1, ayw2, ayw3, ayw4, ayr, adw2, adw4 and adrq+ or adrq-. A study of common HBsAg subtypes in 44 HBsAg reactive sera in University Hospital was conducted using a solid-phase sandwich EIA. Eleven samples were found not typable and among the 33 typable HBsAg reactive sera, 3 HBsAg subtypes: adw, adr and ayw were identified. Subtype adw was found in 66.7% (22/33) of the typable HBsAg reactive sera; 24.2% (8/33) was of subtype adr and 6.0% (2/33) of subtype ayw. One sample was found to be reactive to both adw and adr. HBsAg subtype adw was found more commonly in Chinese but among the Malays, HBsAg subtype adr appeared to predominate. However, the small sample size precludes firm conclusions on the predominant subtype among the Malays.

Matched MeSH terms: Hepatitis B Surface Antigens/analysis*
Fulltext The prevalence of hepatitis B surface antigen and anti-HCV antibody in paediatric oncology patients in Hospital Universiti Sains Malaysia

Menon BS, Aiyar S

Med. J. Malaysia, 1997 Dec;52(4):331-4.
PMID: 10968109

This study examined the prevalence of hepatitis B and C markers in 55 paediatric oncology patients who had completed treatment at the Hospital Universiti Sains Malaysia in Kota Baru. All these children had received blood products and had been treated between 1985-1996. Forty seven per cent of patients were positive for hepatitis B or C. Twenty nine per cent were positive for hepatitis C and twenty two per cent were HBsAg positive. Two children were positive for both and none were HIV positive. Four children had an elevated ALT level and one child had jaundice and hepatomegaly. Some children were marker-positive despite immunization and screening of blood.

Matched MeSH terms: Hepatitis B Surface Antigens/analysis*
Fulltext Sero-prevalence of hepatitis B infection among dental professionals

Vadivale M, Tan TC, Ong CN

Singapore Med J, 1992 Aug;33(4):367-9.
PMID: 1411666

Dental employees in government institutions in a State in Peninsular Malaysia were screened for exposure to hepatitis B virus (HBV) in 1989. Almost all (96.8%) of the 217 employees responded. One quarter (24.8%) was positive for at least one serological markers to HBV; 2.4% had hepatitis B surface antigen (HBsAg) and 22.4% had anti-body to HBsAg (anti-HBs). The presence of HBsAg was unrelated to age, sex, ethnicity, geographical locality and occupations of the subjects. The prevalence of anti-HBs increased with age and was highest for ethnic Chinese (53.6%), followed by Indians (25%), compared to Malays (14.9%) (p less than 0.001) and were increased among dentists (53.1%) and assistant nurses (33.3%). The overall prevalence of HBsAg and anti-HBs were similar to the situation in the community. However, dentists and their chairside assistant nurses, with a higher proportion of Chinese, had higher anti-HBs prevalences compared with that of the general population.

Matched MeSH terms: Hepatitis B Surface Antigens/analysis*
Incidence of hepatitis B infection in Brunei Darussalam--analysis of racial distribution

Alexander MJ, Sinnatamby AS, Rohaimah MJ, Harun AH, Ng JS

Ann. Acad. Med. Singap., 1990 May;19(3):344-6.
PMID: 2393233

Brunei Darussalam has a mixed population with entirely different cultures and religions. The overall incidence of Hepatitis B virus (HBV) infection is 6%. A racial analysis of the incidence of HBV infection in Brunei shows a significantly higher incidence in Chinese compared to the other races. This is consistent with the incidence in the neighbouring countries.

Matched MeSH terms: Hepatitis B Surface Antigens/analysis
Childhood primary hepatocellular carcinoma and hepatitis B virus infection

Cheah PL, Looi LM, Lin HP, Yap SF

Cancer, 1990 Jan 1;65(1):174-6.
PMID: 2152851

In the 7-year period between 1980 and 1987, six cases of childhood primary hepatocellular carcinoma (PHC) were confirmed histologically in our institution. Hepatitis B surface antigen (HBsAg) seropositivity was confirmed in five of the cases, and tissue HBsAg was shown in four of these using the Shikata's orcein stain. An associated maternal HBsAg seropositivity was shown in two of the seropositive children. The youngest seropositive patient who developed PHC was 7 years old. The mother of this patient was also seropositive. These observations support a causal relation between childhood Hepatitis B virus infection and PHC. The importance of vertical or perinatal transmission of HBV in the causation of childhood PHC and the prophylactic role of childhood vaccination is emphasized. Attention is also drawn to the relative short malignant transformation time seen in some of these patients.

Matched MeSH terms: Hepatitis B Surface Antigens/analysis
Fulltext Hepatitis B infection in multitransfused thalassaemics

George E, Ilina I, Yasmin AM, George R, Duraisamy G

Med. J. Malaysia, 1988 Dec;43(4):284-7.
PMID: 3241594
Matched MeSH terms: Hepatitis B Surface Antigens/blood
Fulltext Epidemiological features of hepatitis B virus infection in Malaysians

Ross IN, Dass PK, Thavarasah AS, Noor SS

Med. J. Malaysia, 1988 Dec;43(4):278-83.
PMID: 3241593
Matched MeSH terms: Hepatitis B Surface Antigens/blood
Fulltext Hepatitis B in jaundiced neonates admitted to a special care nursery

Nem-Yun B, Isahak I

Med. J. Malaysia, 1988 Jun;43(2):162-5.
PMID: 3237132
Matched MeSH terms: Hepatitis B Surface Antigens/analysis

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