HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Parhami-Seren, B. Articles by Haupert, G. T. Search for Related Content PubMed PubMed Citation Articles by Parhami-Seren, B. Articles by Haupert, G. T., Jr. Pubmed/NCBI databases Compound via MeSH Substance via MeSH Hazardous Substances DB DIGOXIN OUABAIN

Monoclonal Antibodies That Distinguish Between Two Related Digitalis Glycosides, Ouabain and Digoxin¹

Behnaz Parhami-Seren^*, Charles Bell^2,, Michael N. Margolies^* and Garner T. Haupert, Jr.^3,

^* Department of Surgery, Massachusetts General Hospital and Harvard Medical School, and Renal Unit, Medical Services, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA 02129

The exogenous digitalis glycosides, ouabain and digoxin, have been widely used in humans to treat congestive heart failure and cardiac arrhythmias. Several reports have also pointed to the existence of endogenous ouabain- and digoxin-like compounds, but their precise roles in mammalian physiology and various disorders of the circulation are not clear. In an attempt to produce specific Abs for the purification and identification of endogenous ouabain-like compounds, somatic cell fusion was used to produce mAbs specific for ouabain. Our attempts to produce ouabain-specific mAbs were unsuccessful when ouabain was coupled to exogenous proteins such as bovine -globulins, BSA, and human serum albumin. However, when ouabain was coupled to an Ab of A/J mice origin and the same strain of mouse was used for immunization with ouabain-Ab conjugate, three Abs (1-10, 5A12, and 7-1) specific for ouabain were obtained. In assays of fluorescence quenching and saturation equilibrium with tritiated ouabain, Ab 1-10 exhibited 200 nM affinity for ouabain. These three mAbs are distinguished from existing Abs to ouabain and digoxin by their specificity for ouabain and lack of cross-reactivity with digoxin. Specificity studies showed that the loss of cross-reactivity was correlated with the presence of a hydroxyl group at either position 12ß (digoxin) or 16ß (gitoxin) of the steroid ring. These Abs can be used to develop assays for detection and characterization of ouabain-like molecules in vivo.

This article has been cited by other articles:

				J. R. Murrell, J. D. Randall, J. Rosoff, J.-l. Zhao, R. V. Jensen, S. R. Gullans, and G. T. Haupert Jr Endogenous Ouabain: Upregulation of Steroidogenic Genes in Hypertensive Hypothalamus but Not Adrenal Circulation, August 30, 2005; 112(9): 1301 - 1308. [Abstract] [Full Text] [PDF]

				B. Parhami-Seren, R. Haberly, M. N. Margolies, and G. T. Haupert Jr Ouabain-Binding Protein(s) From Human Plasma Hypertension, August 1, 2002; 40(2): 220 - 228. [Abstract] [Full Text] [PDF]