Summary
The chromatographic behavior of human amniotic interferon on various affinity chromatography ligands was studied. Most of this interferon bound strongly to bovine plasma albumin-agarose, cibacron blue F3 GA-agarose, concanavalin A-agarose and L-tryptophyl-L-tyrosine-θ-carboxyl-pentyl-agarose. After binding most of the interferon activity was eluted only with 50 percent ethylene glycol, showing the high hydrophobicity of this interferon. Smaller quantities could be recovered after phosphate-buffered saline elution or with increased salt concentration. On BPA-θ-carboxy-pentyl-agarose and θ-aminohexyl-agarose, the majority of the biological activity was found in the break-through fraction (eluted with phosphate buffered saline) while some interferon was displaced with high salt or ethylene glycol. Increasing the salt concentration and lowering the pH was necessary to elute interferon from zinc chelate-agarose.
These patterns indicate that human amniotic interferon is similar to human fibroblast (β) interferon but different from human leukocyte (α) interferon. However, the heterogeneity displayed by amniotic interferon on bovine plasma albumin-agarose requires further investigation.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- Am interferon:
-
human amniotic membrane interferon
- BPA:
-
bovine plasma albumin
- CPE:
-
cytopathic effect
- BPA-CH-Sepharose 4B:
-
BPA-θ-carboxyl-pentyl-agarose
- AH-Sepharose 4B:
-
θ-amino-hexyl-agarose
- Con-A-Sepharose-4B:
-
concanavalin A-agarose
- Blue Sepharose CL-6 B:
-
Cibacron Blue F3GA-agarose
- E0 :
-
0.02M phosphate, pH 7.4, 0.15M NaCl
- E1 :
-
0.02M phosphate, pH 7.4, 1M NaCl
- E2 :
-
E1 plus 50 percent ethylene glycol
References
Chen, J. K., Jankowski, W. J., O'Malley, J. A., Sulkowski, E., Carter, W. A.: Nature of the molecular heterogeneity of human leukocyte interferon. J. Virol.19, 425–434 (1976).
Dahl, J., Degre, M.: Separation of antiviral activity of human interferon from cell growth inhibitory effect. Nature257, 799–801 (1975).
Davey, M. J., Huang, J. W., Sulkowski, E., Carter, W. A.: Hydrophobic interaction of human interferon with concanavalin A-agarose. J. biol. Chem.249, 6354–6355 (1974).
Davey, M. J., Huang, J. W., Sulkowski, E., Carter, W. A.: Hydrophobic binding sites on human interferon. J. biol. Chem.250, 348–349 (1975).
Davey, M. W., Sulkowski, E., Carter, W. A.: Binding of human fibroblast interferon to concanavalin A-Agarose. Involvement of carbohydrate recognition and hydrophobic interaction. J. biol. Chem.251, 5381–5385 (1976)
Davey, M. W., Sulkowski, E., Carter, W. A.: Hydrophobic interaction of human, mouse and rabbit interferon with immobilized hydrocarbons. J. biol. Chem.251, 7620–7625 (1976).
Duc-Goiran, P., Galliot, B., Chany, C.: Studies on virus-induced interferons produced by the human amniotic membrane and white blood cells. Arch. ges. Virusforsch.34, 232–243 (1971).
Edy, V. G., Billiau, A., Sommer, P.: Purification of human fibroblast interferon by zinc chelate affinity chromatography. J. biol. Chem.252, 5934–5935 (1977).
Falcoff, E., Fournier, F., Chany, C.: Étude comparative des poids moléculaires de deux interférons humains produitsin vitro. Ann. Inst. Pasteur111, 241–248 (1966).
Ferreira, P. C. P., Peixoto, M. L. P., Silva, M. A. V., Golgher, R. R.: Assay of human interferon in Vero cells by several methods. J. clin. Microbiol.9, 471–475 (1978).
Fournier, F., Falcoff, E., Chany, C.: Demonstration, mass production and characterization of heavy molecular weight human interferon. J. Immunol.99, 1036–1041 (1967).
Grob, P. M., Chadha, K. C.: Separation of human leukocyte interferon components by concanavalin A-agarose affinity chromatography and their characterization. Biochem.18, 5782–5786 (1979).
Havell, E. A., Hayes, T. G., Vilček, J.: Synthesis of two distinct interferons by human fibroblasts. Virology89, 330–334 (1978).
Havell, E. A., Yip, Y. K., Vilček, J.: Characteristics of human lymphoblastoid (Namalva) interferon. J. gen. Virol.38, 51–59 (1977).
Huang, J. W., Davey, M. W., Hejna, C. J.:Muenchhausen, W., Sulkowski, E., Carter, W. A.: Active binding of human interferon albumin immobilized on agarose. J. biol. Chem.249, 4665–4667 (1974).
Huang, J. W., Hejna, C. I., Sulkowski, E., Carter, W. A., Silver, G. H., Munayyer, H., Came, P.: The human interferon-albumin interaction: The influence of albumin concentration. Virology65, 268–271 (1975).
Jankowski, W. J., Davey, M. W., O'Malley, J. A., Sulkowski, E., Carter, W. A.: Molecular structure of human fibroblast and leukocyte interferons: Probe by lectin and hydrophobic chromatography. J. Virol.16, 1124–1130 (1975).
Jankowski, W. J., Muenchhausen, W., Sulkowski, E., Carter, W. A.: Binding of human interferons to immobilized Cibacron Blue F 3 GA: The nature of molecular interaction. Biochem.15, 5182–5187 (1976).
Paucker, K., Dalton, B. J., Ogburn, C. A., Torma, E.: Multiple active sites on human interferons. Proc. Nat. Acad. Sci. (Wash.)72, 4587–4591 (1972).
Pitha, P. M., Marshall, L. W., Carter, W. A.: Interferon: The dissociation of rIn.rCn-induced proteins by protonation. J. gen. Virol.21, 169–174 (1973).
Sulkowski, E., Davey, M. W., Carter, W. A.: Interaction of human interferons with immunobilized hydrophobic amino acids and dipeptides. J. biol. Chem.251, 5381–5385 (1976).
Author information
Authors and Affiliations
Additional information
With 4 Figures
Deceased.
Rights and permissions
About this article
Cite this article
Ferreira, P.C.P., Paucker, M., Golgher, R.R. et al. Affinity chromatography of primary human amnion interferon. Archives of Virology 68, 27–33 (1981). https://doi.org/10.1007/BF01315164
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01315164