Skip to main content
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems
  • Log in
  • My Cart

Main menu

  • Home
  • Articles
    • Archive
  • About the Journal
    • About CVI
    • For Librarians
    • For Advertisers
    • FAQ
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems

User menu

  • Log in
  • My Cart

Search

  • Advanced search
Clinical and Vaccine Immunology
publisher-logosite-logo

Advanced Search

  • Home
  • Articles
    • Archive
  • About the Journal
    • About CVI
    • For Librarians
    • For Advertisers
    • FAQ
CLINICAL LABORATORY IMMUNOLOGY

Development and Technical and Clinical Validation of a Quantitative Enzyme-Linked Immunosorbent Assay for the Detection of Human Antibodies to Hepatitis B Surface Antigen in Recipients of Recombinant Hepatitis B Virus Vaccine

Pierre Cambron, Jeanne-Marie Jacquet, Bernard Hoet, Marc Lievens
Pierre Cambron
GlaxoSmithKline Biologicals, Rue de l'Institut 89, Rixensart, Belgium
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: pierre.A.cambron@gskbio.com
Jeanne-Marie Jacquet
GlaxoSmithKline Biologicals, Rue de l'Institut 89, Rixensart, Belgium
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Bernard Hoet
GlaxoSmithKline Biologicals, Rue de l'Institut 89, Rixensart, Belgium
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Marc Lievens
GlaxoSmithKline Biologicals, Rue de l'Institut 89, Rixensart, Belgium
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
DOI: 10.1128/CVI.00431-08
  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

Article Figures & Data

Figures

  • Tables
  • FIG. 1.
    • Open in new tab
    • Download powerpoint
    FIG. 1.

    Linear regression between FBS and human-plasma anti-HBs free as diluent. Hum-plasma, human plasma.

  • FIG. 2.
    • Open in new tab
    • Download powerpoint
    FIG. 2.

    Observed values and the model predictions for linearity assessment. For groups 1 and 4, the prediction line for each sample is superimposed with the prediction line for the mean sample within each group.

  • FIG. 3.
    • Open in new tab
    • Download powerpoint
    FIG. 3.

    OD response of serial dilutions of three highly concentrated anti-HBs samples.

  • FIG. 4.
    • Open in new tab
    • Download powerpoint
    FIG. 4.

    Deming's regression of the anti-HBs in-house assay and AUSAB RIA with CVs of 11.86% for both the RIA and anti-HBs in-house assay. UL, upper limit; LL, lower limit.

  • FIG. 5.
    • Open in new tab
    • Download powerpoint
    FIG. 5.

    GMCs (mIU/ml) at each blood sampling time point in study A (elderly patients), study E (adolescents), and study F (infants) for the anti-HBs in-house assay versus reference assays. PI to PIV repesent postdose months 1 to 4, respectively, and m1, m2, etc., represent study months 1, 2, etc.

Tables

  • Figures
  • TABLE 1.

    Clinical studies retested with the anti-HBs in-house assay

    StudyPopulationVaccine groupaReference assayNo. of patients in group to be testedTime point(s)bRationale
    A50-70 yr oldExperimental HepB formulationRIA72 (subset)PI, mo 1Verify concordance with AUSAB RIA in an elderly population which reaches lower seroprotection rate and GMT than in adults 18-40 yr old
    Engerix-BPI, mo 1
    PII, mo 6
    PIII, mo 7
    BAdult nonresponderscExperimental HepB formulationRIA58 (all)PreVerify concordance with AUSAB RIA in a difficult population and check specificity of anti-HBs in-house assay on prevaccination samples
    Engerix-BPI, mo 1
    PII, mo 2
    PII, mo 6
    PIII, mo 7
    C11-18 yr oldEngerix-BRIA72 (subset)PI, mo 1Verify concordance with AUSAB RIA after vaccination with a vaccine from another manufacturer
    RecombivaxdPII, mo 2
    PII, mo 6
    PIII, mo 7
    D18-40 yr oldEngerix old 1EIA72 (subset)PII, mo 6Verify concordance with AUSAB EIA following different formulations of the same vaccine
    Engerix old 2PIII, mo 7
    Engerix
    E12-15 yr oldAmbirix 720/20RIA, EIA140 (all)PIII, mo 7Verify concordance with AUSAB RIA and AUSAB EIA
    Twinrix 360/10
    FChildren <1 yr oldInfanrix hexaRIA50 (subset)PreVerify concordance with AUSAB RIA after vaccination with HBV in combined pediatric vaccines
    Infanrix penta + HiberixPII, mo 6
    PII, mo 8
    PIII, mo 9
    GChildren <1 yr oldInfanrix hexaEIA62 (all)Pre, mo 1.5Verify concordance with AUSAB EIA after vaccination with HBV in combined pediatric vaccines
    Infanrix penta + HiberixPIII, mo 7
    PIII, mo 15
    PIV, mo 6
    • ↵ a Engerix old 1, previous formulation, containing thiomersal as a preservative; Engerix old 2, previous formulation, using thiomersal during the production process.

    • ↵ b Pre, sample taken prior to vaccination; PI to PIV, postdose months 1 to 4; mo 1, mo 2, etc., study month 1, study month 2, etc.

    • ↵ c Nonresponders are healthy adults with an anti-HBs antibody concentration of <10 mIU/ml 2 to 5 months after having received at least four doses of HBV.

    • ↵ d Recombivax is a trademark of Merck & Co.

  • TABLE 2.

    Calibration curvea

    Antibody concn (mIU/ml)0.95th percentile
    2.9214.41
    5.848.59
    11.696.31
    23.384.51
    46.754.11
    93.503.12
    187.001.60
    • ↵ a The calibration curve is based on the distribution of the percentage of the absolute values of the relative bias (RE) for the complete set of standard points (0.95 percentile), where % RE = [(recalculated value − assigned value)/assigned value] × 100%.

  • TABLE 3.

    Accuracy of anti-HBs in-house assay in terms of human anti-HBV immunoglobulina

    Theoretical antibody concn (mIU/ml)Measured antibody concn (mIU/ml)% Recovery
    3.33.4100.4
    6.35.690.4
    6.35.893.4
    10.49.894.2
    10.49.490.3
    20.819.995.5
    20.818.990.8
    41.542.0101.2
    41.541.7100.4
    82.681.198.2
    82.680.196.9
    163.9129.979.3
    163.9150.091.5
    476.2446.093.7
    909.1840.692.5
    2,000.01,967.998.4
    3,333.33,304.499.1
    5,000.05,048.4101.0
    10,000.09,885.298.9
    • ↵ a The geometric mean recovery was 94.9%. The 95% CI lower and upper limits were 92.2% and 97.6%, respectively.

  • TABLE 4.

    Demonstration of assay linearitya

    GroupSlope95% CI
    Lower limitUpper limit
    1−0.9990−1.0318−0.9662
    2−0.9910−1.0348−0.9472
    3−0.9802−1.0180−0.9423
    4−0.9976−1.0722−0.9230
    • ↵ a Ten samples with concentrations between 100 and 51,895 mIU/ml were grouped into four groups according to the predilution applied to reach the upper limit of the analytical range. Shown are the estimated slope for each predilution group with its 95% CI using a mixed model with log dilution, group, and samples as factors.

  • TABLE 5.

    Specificity of the anti-HBs in-house assay compared to the AUSAB EIA

    AUSAB EIA antibody concn (mIU/ml) before preincubationAnti-HBs in-house assay antibody concn (mIU/ml) before preincubationHBsAg preincubation with subtype(s)a:
    ad + ayadw2
    AUSAB EIAAnti-HBs in-house assayAUSAB EIAAnti-HBs in-house assay
    Antibody concn (mIU/ml)% SpecificityAntibody concn (mIU/ml)% SpecificityAntibody concn (mIU/ml)% SpecificityAntibody concn (mIU/ml)% Specificity
    2015<3.3100<3.3100<3.3100<3.3100
    9461<3.3100<3.3100<3.3100<3.3100
    119125<3.3100<3.3100<3.3100<3.3100
    346352<3.3100<3.3100<3.3100<3.3100
    382428<3.3100<3.3100<3.3100<3.3100
    1,5561,111<3.3100<3.3100<3.3100<3.3100
    1,9572,410<3.3100<3.3100<3.3100<3.3100
    2,2372,575<3.3100<3.3100<3.3100<3.3100
    49,68042,660<3.3100<3.31008699.84999.9
    54,49061,065<3.3100<3.310065198.810399.8
    <3.3<3.3<3.3NA<3.3NA<3.3<3.3
    <3.3<3.3<3.3NA<3.3NA<3.3<3.3
    <3.3<3.3<3.3NA<3.3NA<3.3<3.3
    • ↵ a Preincubation was performed with either an excess of a mix of native ad and ay HBsAg subtypes or adw2 HBsAg subtype for 2 h at 37°C. Specificity represents the difference in concentrations observed before and after inhibition expressed as a percentage of the corresponding concentration measured without inhibition.

  • TABLE 6.

    Calculation of the average concentration and CV in three samples with high anti-HBs concentrations

    SampleDilutionODAnti-HBs antibody concn (mUI/ml)Avg anti-HBs antibody concn (mUI/ml)aCV (%)
    11>3.000>187
    10,0001.652772,731754,4113
    100,0000.200736,091
    21>3.000>187
    10,0001.438644,921649,4091
    100,0000.180654,897
    31>3.000>187
    10,0001.705833,075823,9762
    100,0000.215814,878
    • ↵ a Average concentration determined at dilutions of 10,000 and 100,000.

  • TABLE 7.

    Contingency table between anti-HBs in-house assay and the AUSAB RIA with different cutoffsa

    Antibody concn cutoff (mIU/ml)Agreement (95% CI)Sensitivity (95% CI)Specificity (95% CI)KappaMcNemar P valueb
    297.71 (96.53-98.58)99.02 (98.08-99.58)87.13 (79.00-92.96)0.881 (0.830-0.931)0.3833
    2.697.82 (96.66-98.67)98.53 (97.45-99.24)92.08 (84.99-96.52)0.891 (0.843-0.938)0.5034
    3.397.61 (96.40-98.49)98.04 (96.84-98.88)94.06 (87.52-97.79)0.883 (0.835-0.931)0.0525
    3.996.74 (95.37-97.79)96.94 (95.52-98.01)95.05 (88.82-98.37)0.847 (0.793-0.900)0.0003
    • ↵ a n = 919.

    • ↵ b Test of dissymmetry in concordance.

  • TABLE 8.

    Bioequivalence of the slope for the anti-HBs in-house assay compared to the AUSAB RIAa

    Equivalence range lower specificationEquivalence range upper specificationEquivalence
    0.502.00Yes
    0.671.50Yes
    0.801.25Yes
    0.901.11Yes
    • ↵ a Shown are the results of Deming regression (ultrastructural model) with CVs of 11.86% for the RIA and the anti-HBS in-house assay. The regression represents a test of bioequivalence on the slope.

  • TABLE 9.

    Seroprotection rates per time point and study for the anti-HBs in-house assay versus the AUSAB RIA and EIA reference assays

    StudyNo. of subjects/groupaTime pointb% Seroprotection in:
    Vaccine group 1Vaccine group 2Vaccine group 3
    In-house assayRIAEIAIn-house assayRIAEIAIn-house assayEIA
    A72PI, mo 115.320.80.02.7
    72PII, mo 280.681.923.626.4
    72PII, mo 695.893.159.756.9
    72PIII, mo 798.698.681.983.3
    B51PI, mo 147.149.063.663.6
    51PII, mo 260.860.881.881.8
    50PII, mo 648.042.085.281.5
    47PIII, mo 772.372.396.296.2
    C100PI, mo 12.02.09.712.5
    72PII, mo 280.668.168.169.4
    72PII, mo 693.193.195.895.8
    72PIII, mo 798.697.2100.0100.0
    D72PIII, mo 797.295.897.297.297.297.2
    E134PI/II, mo 253.040.341.890.486.886.8
    145PII/III, mo 799.398.699.3100.0100.0100.0
    F50Pre13.816.912.212.2
    50PII, mo 698.098.096.096.0
    50PII, mo 894.094.0100.0100.0
    50PIII, mo 9100.0100.0100.0100.0
    G34Pre, mo 1.552.949.059.668.4
    47PIII, mo 7100.0100.095.995.0
    53PIII, mo 1596.696.693.096.0
    57PIV, mo 6100.0100.098.398.3
    • ↵ a If the number between the two groups was different, the lowest number was taken.

    • ↵ b Pre, sample taken prior to vaccination; PI to PIV, postdose months 1 to 4, respectively; mo 1, mo 2, etc., study month 1, study month 2, etc.

PreviousNext
Back to top
Download PDF
Citation Tools
Development and Technical and Clinical Validation of a Quantitative Enzyme-Linked Immunosorbent Assay for the Detection of Human Antibodies to Hepatitis B Surface Antigen in Recipients of Recombinant Hepatitis B Virus Vaccine
Pierre Cambron, Jeanne-Marie Jacquet, Bernard Hoet, Marc Lievens
Clinical and Vaccine Immunology Jul 2009, 16 (8) 1236-1246; DOI: 10.1128/CVI.00431-08

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Print

Email

Thank you for sharing this Clinical and Vaccine Immunology article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
Development and Technical and Clinical Validation of a Quantitative Enzyme-Linked Immunosorbent Assay for the Detection of Human Antibodies to Hepatitis B Surface Antigen in Recipients of Recombinant Hepatitis B Virus Vaccine
(Your Name) has forwarded a page to you from Clinical and Vaccine Immunology
(Your Name) thought you would be interested in this article in Clinical and Vaccine Immunology.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Development and Technical and Clinical Validation of a Quantitative Enzyme-Linked Immunosorbent Assay for the Detection of Human Antibodies to Hepatitis B Surface Antigen in Recipients of Recombinant Hepatitis B Virus Vaccine
Pierre Cambron, Jeanne-Marie Jacquet, Bernard Hoet, Marc Lievens
Clinical and Vaccine Immunology Jul 2009, 16 (8) 1236-1246; DOI: 10.1128/CVI.00431-08
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Top
  • Article
    • ABSTRACT
    • MATERIALS AND METHODS
    • RESULTS
    • DISCUSSION
    • ACKNOWLEDGMENTS
    • FOOTNOTES
    • REFERENCES
  • Figures & Data
  • Info & Metrics
  • PDF

KEYWORDS

Enzyme-Linked Immunosorbent Assay
Hepatitis B Antibodies
Hepatitis B Surface Antigens
Hepatitis B Vaccines
Hepatitis B virus

Related Articles

Cited By...

About

  • About CVI
  • For Librarians
  • For Advertisers
  • FAQ
  • Permissions
  • Journal Announcements

Authors

  • Submit a Manuscript to mSphere

ASM Journals

ASM journals are the most prominent publications in the field, delivering up-to-date and authoritative coverage of both basic and clinical microbiology.

About ASM | Contact Us | Press Room

 

ASM is a member of

Scientific Society Publisher Alliance

 

American Society for Microbiology
1752 N St. NW
Washington, DC 20036
Phone: (202) 737-3600

Copyright © 2021 American Society for Microbiology | Privacy Policy | Website feedback

Print ISSN: 1556-6811; Online ISSN: 1556-679X