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Use of an Aqueous Polymer Phase System as an Assay for Antibodies to Escherichia Coli Alkaline Phosphatase¹

From the Department of Microbiology, Yale University, New Haven, Connecticut 06510

Abstract

Escherichia coli alkaline phosphatase in both crude and purified forms was used to produce antibody in rabbits. With this antigen-antibody system it was demonstrated that a polyethyleneglycol-dextran aqueous polymer phase system could be used as a rapid, sensitive and precise means of measuring quantitatively the interaction of antibodies and biologically active macromolecules. Similar results were obtained whether crude or highly pure forms of the enzyme were used. In experiments using nanogram quantities of both antigen (190 ng) and antibody (27 to 32 ng), it was possible to demonstrate differences between the three electrophoretic forms of this enzyme which are thought to be coded for by a single structural gene. Serologic phase system analysis was shown to be versatile enough to compare the native enzyme with forms of the enzyme which were virtually inactive (CRM) as well as with the Zn(II) and Co(II) reconstituted apoenzyme.

Footnotes

¹ This work was supported by United States Public Health Service Training Grant AI00291.

² Present address: Department of Biology, Johns Hopkins University, Baltimore, Maryland 21218.