Immunological and biophysical properties of hepatitis B antigen labeled by the chloramine-T and by the lactoperoxidase methods

Hollinger, F. Blaine; Morrison, Martin; Chairez, Ruben; Dreesman, Gordon R.

doi:10.1016/0022-1759(75)90083-6

Cited by 16 publications

(6 citation statements)

References 23 publications

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“…Iodination of HBcAg and various other proteins with 125I was carried out by the chloramine-T method of G reenwood et al [12], with modifications described by Hollinger et al [13]. Purified preparations of HBcAg (0.1 ml) were iodinated and either used directly for PAGE or desalted in Sepharose CL-4B.…”

Section: Methodsmentioning

confidence: 99%

Biochemical and Biophysical Properties of Hepatitis B Core Particles Derived from Dane Particles and Infected Hepatocytes

Desmyter

et al. 1977

Intervirology

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Hepatitis B core antigen (HBcAg) was purified from Dane particles and from infected hepatocytes. An identical isoelectric pH of 4.0 was determined for labeled preparations of both Dane-derived and liver-derived HBcAg. Unlabeled liver-derived HBcAg demonstrated a lower isoelectric pH of 3.7. Molecular weight determinations by Sepharose 4B column chromatography revealed that liver-derived HBcAg had a molecular weight of 8.5–9.0 × 106 daltons. The sedimentation coefficient of both Dane- and liver-derived HBcAg was found to be 124S. PAGE revealed that iodinated HBcAg derived from Dane particles was very similar in polypeptide structure to HBcAg derived from infected liver tissue. Twelve polypeptides were resolved from Dane core particles, and seven to nine were resolved from liver core particles. Several of the polypeptides in both preparations co-migrated with iodinated hepatitis B surface antigen (HBsAg). However, three polypeptides (mol. wt. 88,000, 79,000 and 59,000) were found in both Dane- and liver-derived HBcAg but not in HBsAg, which suggests that these polypeptides are HBcAg-specific. Endogenous DNA polymerase activity was observed in both Dane- and liver-derived core particles.

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Section: Methodsmentioning

confidence: 99%

Biochemical and Biophysical Properties of Hepatitis B Core Particles Derived from Dane Particles and Infected Hepatocytes

Desmyter

et al. 1977

Intervirology

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“…act. 7-12 pCi/pg) [8,9] in 75 pi of PBS containing 25% FCS was added. The human IgG was purified by DEAE-cellulose chromatography from plasma with an anti-HBc complement-fixing titer of 1:1,024.…”

Section: Methodsmentioning

confidence: 99%

Endogenous DNA Polymerase-Positive Core Particles from Hepatitis-Infected Hepatocytes

Ha¹,

Gr²,

Fb³

et al. 1975

Intervirology

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The purification is described of liver-derived hepatitis B core antigen (HBe Ag) from human hepatocytes demonstrating only intranuclear particles by electron microscopy. The purified preparation contained 5 × 10u particles per ml. The particles were mono-dispersed and relatively free of background material. DNA-dependent polymerase activity was present and had greater activity than an equal number of cores derived from Dane particles isolated from plasma. Specificity of the polymerase reaction was confirmed by precipitation of the activity with specific anti-HBc antiserum. A proportion of the liver-derived core particles was nonreactive for DNA polymerase activity. The polymerase-positive population of particles had a larger size than the polymerase-negative population of liver-derived cores as evidenced by gel filtration in Sepharose 4B.

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“…More than 90% of the labeled material was found in the cathode solution (pH < 3.0), indicating that the antigen had been extensively oxidized (data not shown). Less than 0.1% of the label was found at a pI of 3.7, where untreated, labeled HBsAg would be expected (4,13).…”

Section: Treatment Of Hbsag-positive Plasma Withmentioning

confidence: 92%

“…In vitro radiolabeling of HBsAg and human anti-HBc with 1'I. Purified preparations of HBsAg (25 to 30 jAg) were labeled with 1 mCi of '251, using the chloramine T method of Greenwood et al (11) as modified by Hollinger and co-workers (13). lodinated HBsAg used in these experiments ranged in specific activity from 10 to 25 ,iCi of "1I per ,g of HBsAg.…”

Section: Treatment Of Hbsag-positive Plasma Withmentioning

confidence: 99%

Immunological and biophysical alteration of hepatitis B virus antigens by sodium hypochlorite disinfection

Sehulster¹,

Hollinger²,

Dreesman³

et al. 1981

Appl Environ Microbiol

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Sodium hypochlorite (NaOCl) was examined as an effective disinfectant in hepatitis laboratories. Concentrations of NaOCl containing 5,600 ppm (5,600 ,ug/ ml) of available chlorine were found to be effective in destroying the antigenicity of hepatitis B surface antigen (HBsAg) in virion-rich plasma after an exposure time of 1 min or more. In the treatment of protein-deficient solutions containing HBsAg, smaller concentrations of available chlorine (<500 ppm) are equally effective. Neither 17to 25-nm HBsAg particles nor 45-nm virion particles could be detected by electron microscopy after treatment. Chemical interaction of protein and NaOCl was confirmed by isoelectrofocusing of 125I-labeled HBsAg. More than 90% of the labeled material was found at pH 3.0 or lower, indicating complete antigen oxidation. Labeled HBsAg was reduced in density from 1.21 g/ cm3 in CsCl to approximately 1.07 g/cm3 after treatment with NaOCl. Both hepatitis B core antigen and deoxyribonucleic acid polymerase activity were significantly reduced after interaction with hypochlorite solutions. These results show that NaOCl destroys hepatitis B antigenicity and virus structures and therefore may be utilized as a disinfectant for the virus.

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Immunological and biophysical properties of hepatitis B antigen labeled by the chloramine-T and by the lactoperoxidase methods

Cited by 16 publications

References 23 publications

Biochemical and Biophysical Properties of Hepatitis B Core Particles Derived from Dane Particles and Infected Hepatocytes

Biochemical and Biophysical Properties of Hepatitis B Core Particles Derived from Dane Particles and Infected Hepatocytes

Endogenous DNA Polymerase-Positive Core Particles from Hepatitis-Infected Hepatocytes

Immunological and biophysical alteration of hepatitis B virus antigens by sodium hypochlorite disinfection

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