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Home / Equine Research Bank / Appl Microbiol / Preparation and standardization of an Australia antigen anti...
Applied microbiology1971; 21(6); 1017-1023; doi: 10.1128/am.21.6.1017-1023.1971

Preparation and standardization of an Australia antigen antibody of equine origin.

Abstract: A horse has been immunized with Australia antigen (Au/SH) purified 20-fold by a procedure employing gel filtration of Cohn fraction IV derived from an Au/SH-positive human plasma pool. Hyperimmunization was initiated by the intramuscular injection of 20 ml of a mixture of equal parts of purified Au/SH and complete Freund's adjuvant. The 20-ml volume was divided into four 5-ml doses, two of which were administered on each side of the horse's neck. Booster doses of antigen alone were given as follows: 10 ml intravenously 30 days later and 5 ml intramuscularly on each of days 77 and 205. Au/SH antibody formed readily, beginning on day 17, and was demonstrated by the agar gel double-diffusion technique and the complement fixation test during the subsequent 6 months. Antihuman plasma protein antibodies were effectively removed from the horse serum by one absorption with 1 to 3 volumes of normal human plasma. Abrupt rises in anticomplementary activity observed shortly after the third and fourth antigen injections, when the horse had developed elevated and steady levels of Au/SH antibody, could possibly be due to formation of antigen-antibody complexes. After optimal conditions were determined, an Au/SH antibody reagent pool which met official requirements was prepared. It was found equally suitable for the agar gel double-diffusion, complement fixation, and counterimmunoelectrophoresis test procedures.
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Publication Date: 1971-06-01 PubMed ID: 4998346PubMed Central: PMC377335DOI: 10.1128/am.21.6.1017-1023.1971Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The study presents how a horse was immunized with Australia antigen (Au/SH), a vital component in identifying hepatitis B, the resulting antibody production, and subsequent use of these antibodies as a reagent for various diagnostic tests.

Methodology

  • The researchers purified the Australia antigen 20-fold via a procedure involving gel filtration of a DNA component (Cohn fraction IV) acquired from a human plasma pool that tested positive for the antigen (Au/SH).
  • The horse was then immunized via an intramuscular injection of a mixture of equal parts of the purified antigen and a compound (Freund’s adjuvant) that boosts immune response.
  • The initial dose was divided into four equal parts and injected in both sides of the horse’s neck. The horse was subsequently given booster doses of the antigen alone at specific time intervals (on the 30th day, then on the 77th day, and the 205th day).

Results

  • Antibodies against the antigen started forming by day 17, and were detectable over the next six months through techniques such as the agar gel double-diffusion and the complement fixation test.
  • Any antibodies created against human proteins present in the horse’s serum, a potentially interfering factor, were effectively removed by treating the serum with normal human plasma.
  • Brief surges in the activity against complement proteins, usually involved in immune responses, were observed following the third and fourth antigen shots. This change could be a result of the formation of antigen-antibody complexes.

Utilization of Antibodies

  • After optimal conditions were determined, a pool of the Australia antigen antibody reagent was created. This reagent met official requirements and was deemed suitable for use in various diagnostic procedures such as the agar gel double-diffusion, complement fixation, and counterimmunoelectrophoresis test procedures.

This research provides important data on creating a reliable source of hepatitis B virus (HBV) antibodies, which can be utilized in different diagnostic tests. This could improve the accuracy and speed of HBV diagnosis, leading to more efficient patient treatment.

Cite This Article

APA
Cabasso VJ, Nieman R, Schroeder DD, Hok KA, Louie RE, Mozen MM. (1971). Preparation and standardization of an Australia antigen antibody of equine origin. Appl Microbiol, 21(6), 1017-1023. https://doi.org/10.1128/am.21.6.1017-1023.1971

Publication

Applied microbiology
ISSN: 0003-6919
NlmUniqueID: 7605802
Country: United States
Language: English
Volume: 21
Issue: 6
Pages: 1017-1023

Researcher Affiliations

Cabasso, V J
    Nieman, R
      Schroeder, D D
        Hok, K A
          Louie, R E
            Mozen, M M

              MeSH Terms

              • Animals
              • Antibodies / analysis
              • Antibodies / isolation & purification
              • Chromatography, Gel
              • Complement Fixation Tests
              • Hepatitis B Antigens
              • Hepatitis B virus / immunology
              • Hepatitis B virus / isolation & purification
              • Horses
              • Humans
              • Immune Sera
              • Immunization
              • Immunochemistry
              • Immunodiffusion
              • Immunoelectrophoresis
              • Injections, Intramuscular
              • Injections, Intravenous
              • Time Factors

              References

              This article includes 14 references
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                pubmed: 4245401doi: 10.3181/00379727-133-34705google scholar: lookup
              9. Gocke DJ, Howe C. Rapid detection of Australia antigen by counterimmunoelectrophoresis.. J Immunol 1970 Apr;104(4):1031-4.
                pubmed: 4986160
              10. Schroeder DD, Mozen MM. Australia antigen: distribution during Cohn ethanol fractionation of human plasma.. Science 1970 Jun 19;168(3938):1462-4.
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              11. Hok KA, Nieman R, Lackey JO, Cabasso VJ. Australia antigen in a closed adult population monitored for serum glutamic oxalacetic transaminase.. Appl Microbiol 1970 Jul;20(1):6-10.
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              12. Taswell HF, Shorter R, Poncelet TK, Maxwell NG. Hepatitis-associated antigen in blood donor populations. Relationship to posttransfusion hepatitis.. JAMA 1970 Oct 5;214(1):142-4.
                pubmed: 4989996
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              Citations

              This article has been cited 5 times.
              1. Anderson KE, Sun SC, Berg HS, Chang NK. Liver function and histology in asymptomatic Chinese military personnel with hepatitis B antigenemia. Am J Dig Dis 1974 Aug;19(8):693-703.
                doi: 10.1007/BF01844938pubmed: 4843213google scholar: lookup
              2. Cruceanu A, Müller R, Deicher H. A simple procedure for purification of hepatitis B-antigen. Med Microbiol Immunol 1973 Sep 26;159(1):83-8.
                doi: 10.1007/BF02122652pubmed: 4769894google scholar: lookup
              3. Dreesman GR, Hollinger FB, McCombs RM, Melnick JL. Production of potent anti-Australia antigen sera of high specificity and sensitivity in goats. Infect Immun 1972 Feb;5(2):213-21.
                doi: 10.1128/iai.5.2.213-221.1972pubmed: 4635499google scholar: lookup
              4. Shaw ED, McKee AP, Rancourt M, Hollenbeck L. Induction of hepatitis B antibody in experimental animals by immunization with A-2 plaque virus. J Virol 1973 Dec;12(6):1598-607.
                doi: 10.1128/JVI.12.6.1598-1607.1973pubmed: 4128387google scholar: lookup
              5. De Rizzo E, Pandey R, Wallis C, Melnick JL. Concentration and purification of hepatitis B antigen with polyethylene glycol and polyelectrolyte 60, a cross-linked copolymer of isobutylene maleic anhydride. Infect Immun 1972 Sep;6(3):335-8.
                doi: 10.1128/iai.6.3.335-338.1972pubmed: 4118047google scholar: lookup
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