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. 1962 Jan 1;115(1):195–208. doi: 10.1084/jem.115.1.195

PHYSICAL PROPERTIES OF ANTIBODY TO BOVINE SERUM ALBUMIN AS DEMONSTRATED BY HEMAGGLUTINATION

Albert A Benedict 1, Ronald J Brown 1, Rajalakshmi Ayengar 1
PMCID: PMC2137484  PMID: 13867131

Abstract

Rabbit antibovine serum albumin antisera were fractionated by zone electrophoresis on starch, zone ultracentrifugation in sucrose density gradients, and diethylaminoethyl-cellulose chromatography, and were assayed by the passive HA technique. Antisera had at least two antibodies: one associated with a fraction characterized as a γ-2 globulin with a sedimentation rate of 7.3S and low anionic binding (fraction I), and one associated with a fraction characterized as a γ-1 globulin (or β-globulin) with a sedimentation rate of 20.4S and high anionic binding (fraction IV). The HA titers of fractions I and IV of early sera were approximately equal. On prolonged antigenic stimulation fraction IV agglutinin decreased in concentration and the relative concentration of fraction I agglutinin increased. Chromatographic analysis of hyper-immune sera yielded 2 additional HA fractions with intermediate anionic binding. These results indicated that rabbits synthesize anti-BSA antibodies similar to rabbit anti-cellular antibodies. Rabbit anti-BSA precipitin migrated mainly as a γ-2 globulin and the agglutinin migrated with γ- and β-globulins. The evidence suggested that the HA method might measure antibody not measured by the quantitative precipitin technique.

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Selected References

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

  1. ABELSON N. M., RAWSON A. J. [Studies of blood group antibodies. I. Fractionation of anti-A and anti-B Isohemagglutinins by anioncation cellulose exchange chromatography]. J Immunol. 1959 May;82(5):435–443. [PubMed] [Google Scholar]
  2. BENEDICT A. A., BRIGGS N. T. Separation of psittacosis complement fixing and hemagglutinating antibodies by starch electrophoresis. Virology. 1958 Dec;6(3):772–773. doi: 10.1016/0042-6822(58)90124-7. [DOI] [PubMed] [Google Scholar]
  3. BOYDEN S. V. The adsorption of proteins on erythrocytes treated with tannic acid and subsequent hemagglutination by antiprotein sera. J Exp Med. 1951 Feb;93(2):107–120. doi: 10.1084/jem.93.2.107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. BRAKKE M. K. Zone electrophoresis of dyes, proteins and viruses in density-gradient columns of sucrose solutions. Arch Biochem Biophys. 1955 Mar;55(1):175–190. doi: 10.1016/0003-9861(55)90556-7. [DOI] [PubMed] [Google Scholar]
  5. CAMPBELL D. H., STURGEON P., VINOGRAD J. R. Separation of complete and incomplete Rh antibodies by centrifugation. Science. 1955 Dec 2;122(3179):1091–1092. doi: 10.1126/science.122.3179.1091-b. [DOI] [PubMed] [Google Scholar]
  6. DIXON F. J., MAURER P. H., DEICHMILLER M. P. Primary and specific anamnestic antibody responses of rabbits to heterologous serum protein antigens. J Immunol. 1954 Feb;72(2):179–186. [PubMed] [Google Scholar]
  7. FUDENBERG H. H., KUNKEL H. G. Physical properties of the red cell agglutinins in acquired hemolytic anemia. J Exp Med. 1957 Nov 1;106(5):689–702. doi: 10.1084/jem.106.5.689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Heidelberger M., Pedersen K. O. THE MOLECULAR WEIGHT OF ANTIBODIES. J Exp Med. 1937 Feb 28;65(3):393–414. doi: 10.1084/jem.65.3.393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. KUNKEL H. G., SLATER R. J. Zone electrophoresis in a starch supporting medium. Proc Soc Exp Biol Med. 1952 May;80(1):42–44. doi: 10.3181/00379727-80-19516. [DOI] [PubMed] [Google Scholar]
  10. Kekwick R. A. The serum proteins in multiple myelomatosis. Biochem J. 1940 Sep;34(8-9):1248–1257. doi: 10.1042/bj0341248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  12. PORTER R. J. Studies on antibody formation. Effects of x-irradiation on adaptation for the secondary response of rabbits to bovine gamma-globulin. J Immunol. 1960 May;84:485–490. [PubMed] [Google Scholar]
  13. PORTER R. R. The fractionation of rabbit gamma-globulin by partition chromatography. Biochem J. 1955 Mar;59(3):405–410. doi: 10.1042/bj0590405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. PRESSMAN D., JAMES A. W., YAGI Y., HIRAMOTO R., WOERNLEY D., MAXWELL W. T. Rapidly sedimenting properties of specifically precipitating component of a Hashimoto's disease serum. Proc Soc Exp Biol Med. 1957 Dec;96(3):773–777. doi: 10.3181/00379727-96-23604. [DOI] [PubMed] [Google Scholar]
  15. SCHEIDEGGER J. J. Une micro-méthode de l'immuno-electrophorèse. Int Arch Allergy Appl Immunol. 1955;7(2):103–110. [PubMed] [Google Scholar]
  16. STAVITSKY A. B. Micromethods for the study of proteins and antibodies. I. Procedure and general applications of hemagglutination and hemagglutination-inhibition reactions with tannic acid and protein-treated red blood cells. J Immunol. 1954 May;72(5):360–367. [PubMed] [Google Scholar]
  17. STELOS P. Comparative study of rabbit hemolysins to various antigens. I. Hemolysins to beef red cells. J Infect Dis. 1958 Mar-Apr;102(2):103–113. doi: 10.1093/infdis/102.2.103. [DOI] [PubMed] [Google Scholar]

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