Sep 1, 1993

A study of the advantages and limitations of immunoblotting procedures for the detection of antibodies against influenza virus

Electrophoresis
G P Kapaklis-DeliyannisD C Jackson

Abstract

An immunoblotting procedure was used to determine the specificity and examine some of the properties of antibodies produced following infection of mice with influenza virus or inoculation with noninfectious material with Alhydrogel or complete Freund's adjuvant. The noninfectious material used was beta-propiolactone-inactivated influenza virus and a preparation (HANA) enriched for the surface glycoproteins, hemagglutinin (HA) and neuraminidase (NA). When influenza viral proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under nonreducing conditions, each of the anti-viral antisera tested exhibited strong binding. Under reducing conditions, however, much weaker binding was observed especially towards the HA1 subunit of HA. This was particularly apparent with antisera raised to virus or HANA in the absence of adjuvant. A panel of monoclonal antibodies directed to HA also bound well to viral HA separated by SDS-PAGE under nonreducing conditions but failed to recognize epitopes on HA1 separated under reducing conditions. These results suggest that when HA is reduced and immobilized on a solid support, it does not display the conformational features essential for the integrity of all epit...Continue Reading

  • References37
  • Citations3

References

Mentioned in this Paper

Monoclonal Antibodies
Monoclonal antibodies, antineoplastic
Immunoblotting, Reverse
Neu1
Antibodies, Viral
Mice, Inbred BALB C
Immunologic Adjuvants
Adjuvant
Virion
SDS-PAGE

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