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Differences in the isoelectric focussing pattern of antibodies to human serum albumin eluted from an immunoadsorbent (HSA-sepharose) with thiocyanate ions

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Research in Experimental Medicine

Summary

Anti HSA-antibodies were isolated from HSA-sepharose by direct and stepwise elution with 0.5 M, 2.0 and 3.0 M KSCN at different pH values. The antibody fractions, especially those eluted stepwise, varied in their isoelectrofocussing pattern. Nearly all antibody molecules with an isoelectric point (PI) lower than 6.5 were eluted with 0.5 M KSCN. Application of increasing KSCN concentrations led to the recovery of antibody fractions showing a parallel shift in the PI to the region higher than 7.5. These differences were not correlated with differences in antibody avidity. The effect of KSCN does not seem to be restricted to the combining sites of the antibody. The dissociation of the antibody from the antigen is apparently facilitated by an interaction of KSCN with the remainder of the antibody molecule thereby influencing the overall net charge.

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References

  1. Dandliker, W. B., Alonso, R., de Saussure, V. A., Kierszenbaum, F., Levison, S. A., Shapiro, H.C.: The effect of chaotropic ions on the dissociation of antigen-antibody complexes. Biochemistry6, 1460 (1967)

    PubMed  Google Scholar 

  2. Dandliker, W. B., de Saussure, V. A., Levandoski, N.: Antibody purification at neutral pH utilizing immunospecific adsorbents. Immunochemistry5, 357 (1968)

    PubMed  Google Scholar 

  3. de Saussure, V. A., Dandliker, W. B.: Ultracentrifuge studies of the effects of thiocyanate ion on antigen-antibody systems. Immunochemistry6, 77 (1969)

    PubMed  Google Scholar 

  4. McConahey, P. J., Dixon, F. J.: A method of trace iodination of proteins for immunologic studies. Int. Arch. Allergy29, 185 (1966)

    PubMed  Google Scholar 

  5. Kist, M., Vogt, A., Heinzel, W.: Isolation of antiferritin antibody. Immunochemistry12, 119 (1975)

    Google Scholar 

  6. Shimizu, F., Mossmann, H., Himmelspach, K., Takamiya, H., Vogt, A.: Separation of antidinitrophenyl antibodies according to affinity by fractionated elution from immunoadsorbent using thiocyanate. (In prep.)

  7. Takamiya, H., Shimizu, F., Vogt, A.: A two stage method for cross-linking antibody globulin to ferritin by glutaraldehyde. II. Antigen binding and agglutinating capacity. J. Immunol. Methods4, 381 (1974)

    PubMed  Google Scholar 

  8. Weiler, I. J., Bosma, G., Dittrich, B., Weiler, E.: Analysis of antibody populations to oligo-D-alanine. VIII. Sequential studies on properties of antibody subpopulations: rate constant, affinity, and isoelectric focusing spectrum. Eur. J. Immunol.5, 399 (1975)

    Google Scholar 

  9. Woehler, M. E., Lovins, R. E.: A comparison of the isoelectrofocusing properties of antibodies to DNP, DNP-glycylglycylglycine and DNP-p-aminobenzoylglutamate. Immunology28, 1089 (1975)

    PubMed  Google Scholar 

  10. Yoo, T. J.: Isoelectric distribution of specifically purified rabbit antibody against 4-azonaphthalene-1-sulfonate. Immunochemistry12, 199 (1975)

    PubMed  Google Scholar 

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Shimizu, F., Kist, M. & Vogt, A. Differences in the isoelectric focussing pattern of antibodies to human serum albumin eluted from an immunoadsorbent (HSA-sepharose) with thiocyanate ions. Res. Exp. Med. 172, 231–238 (1978). https://doi.org/10.1007/BF01855833

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  • DOI: https://doi.org/10.1007/BF01855833

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