Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1973 Oct;8(4):645–649. doi: 10.1128/iai.8.4.645-649.1973

Coupling of Peroxidase to Poliovirus Antibody: Characteristics of the Conjugates and Their Use in Virus Detection

John E Herrmann 1, Stephen A Morse 1
PMCID: PMC422904  PMID: 4126429

Abstract

Antibody to poliovirus type 1 (Po-1) was coupled to peroxidase by use of glutaraldehyde or 4, 4′ -difluoro,3, 3′ -dinitro diphenyl sulfone. Glutaraldehyde was found to be the superior coupling agent, yielding conjugates that had up to 2.8 × 104 enzyme units/ml (75% of total enzyme input). Conjugates migrated as a single band when centrifuged in sucrose density gradients, demonstrating that the purification procedure used was effective in removing both noncoupled enzyme and heterogeneous antibody components. Conjugates were specific for Po-1 and did not adsorb to cells infected with unrelated enterovirus types. Adsorption of conjugates to Po-1-infected cells was demonstrable within 6 h postinfection.

Full text

PDF
645

Images in this article

647Image
on p.647

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abelson H. T., Smith G. H., Hoffman H. A., Rowe W. P. Use of enzyme-labeled antibody for electron microscope localization of lymphocytic choriomeningitis virus antigens in infected cell cultures. J Natl Cancer Inst. 1969 Mar;42(3):497–515. [PubMed] [Google Scholar]
  2. Avrameas S. Coupling of enzymes to proteins with glutaraldehyde. Use of the conjugates for the detection of antigens and antibodies. Immunochemistry. 1969 Jan;6(1):43–52. doi: 10.1016/0019-2791(69)90177-3. [DOI] [PubMed] [Google Scholar]
  3. Dougherty R. M., Marucci A. A., Distefano H. S. Application of immunohistochemistry to study of avian leukosis virus. J Gen Virol. 1972 May;15(2):149–162. doi: 10.1099/0022-1317-15-2-149. [DOI] [PubMed] [Google Scholar]
  4. Kurstak E., Belloncik S., Onji P. A., Montplaisir S., Martineau B. Localisation par l'immunoperoxydase des antigènes du virus cytomégalique en culture cellulaire de fibroblastes humains. Microscopie photonique et électronique. Arch Gesamte Virusforsch. 1972;38(1):67–76. [PubMed] [Google Scholar]
  5. Leduc E. H., Wicker R., Avrameas S., Bernhard W. Ultrastructural localization of SV40T antigen with enzyme-labelled antibody. J Gen Virol. 1969 Jun;4(4):609–614. doi: 10.1099/0022-1317-4-4-609. [DOI] [PubMed] [Google Scholar]
  6. Nakane P K, Pierce G B., Jr Enzyme-labeled antibodies: preparation and application for the localization of antigens. J Histochem Cytochem. 1966 Dec;14(12):929–931. doi: 10.1177/14.12.929. [DOI] [PubMed] [Google Scholar]
  7. ROTMAN B. Measurement of activity of single molecules of beta-D-galactosidase. Proc Natl Acad Sci U S A. 1961 Dec 15;47:1981–1991. doi: 10.1073/pnas.47.12.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Shabo A. L., Petricciani J. C., Kirschstein R. L. Immunoperoxidase localization of herpes zoster virus and simian virus 40 in cell culture. Appl Microbiol. 1972 May;23(5):1001–1009. doi: 10.1128/am.23.5.1001-1009.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ubertini T., Wilkie B. N., Noronha F. Use of horseradish peroxidase-labeled antibody for light and electron microscope localization of reovirus antigen. Appl Microbiol. 1971 Mar;21(3):534–538. doi: 10.1128/am.21.3.534-538.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Wicker R., Avrameas S. Localization of virus antigens by enzyme-labelled antibodies. J Gen Virol. 1969 Jun;4(4):465–471. doi: 10.1099/0022-1317-4-4-465. [DOI] [PubMed] [Google Scholar]
  11. Wicker R. Comparison of immunofluorescent and immunoenzymatic techniques applied to the study of viral antigens. Ann N Y Acad Sci. 1971 Jun 21;177:490–500. doi: 10.1111/j.1749-6632.1971.tb35079.x. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES