Clinical Chemistry AACC Online Job Center
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Clinical Chemistry 35: 985-989, 1989;
This Article
Right arrow Full Text (PDF)
Right arrow Submit an electronic Letter to
the Editor about this paper
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Landt, Y.
Right arrow Articles by Ladenson, J. H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Landt, Y.
Right arrow Articles by Ladenson, J. H.

Clinical Chemistry, Vol 35, 985-989, Copyright © 1989 by American Association for Clinical Chemistry

Immunoaffinity purification of creatine kinase-MB from human, dog, and rabbit heart with use of a monoclonal antibody specific for CK-MB

Y Landt, HC Vaidya, SE Porter, DN Dietzler and JH Ladenson
Department of Pathology, Washington University School of Medicine, St. Louis, MO 63110.

We describe a simple, rapid immunoaffinity procedure for purifying the MB isoenzyme of creatine kinase. Immunoaffinity gel is prepared by linking a monoclonal antibody ("Conan-MB"), specific for this isoenzyme, to Sepharose 4B. Heart tissue is homogenized and fractionated with 40-70% saturated ammonium sulfate before it is applied to the immunoaffinity gel. CK-MB activity, retained on the gel, is then eluted with a high-pH diethylamine buffer (0.1 mol/L, pH 10.5). The purified CK-MB isoenzyme is stabilized by collection directly into tubes containing glycerol (to prevent dissociation of the enzyme subunits) and pH-neutralizing buffer. This procedure compares favorably in yield, specific activity, and technical ease with a multi-column purification method previously used in our laboratory. We have used the immunoaffinity procedure to purify to homogeneity CK-MB from human, dog, and rabbit heart, with yields of 50.0%, 53.1%, and 49.3% and specific activities of 540, 477, and 377 kU/g, respectively. The preparations are pure as judged by protein staining with silver nitrate after electrophoresis on sodium dodecyl sulfate/polyacrylamide gel.


The following articles in journals at HighWire Press have cited this article:


Home page
 Toxicol PatholHome page
D. B. Walker
Serum Chemical Biomarkers of Cardiac Injury for Nonclinical Safety Testing
Toxicol Pathol, January 1, 2006; 34(1): 94 - 104.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Chem.Home page

Clin. Chem., May 1, 2002; 48(5): 805 - 805.
[Full Text] [PDF]

Home page
 Clin. Chem.Home page
R. H. Christenson, H. Vaidya, Y. Landt, R. S. Bauer, S. F. Green, F. A. Apple, A. Jacob, G. R. Magneson, S. Nag, A. H.B. Wu, et al.
Standardization of Creatine Kinase-MB (CK-MB) Mass Assays: The Use of Recombinant CK-MB as a Reference Material
Clin. Chem., September 1, 1999; 45(9): 1414 - 1423.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 1989 by the American Association for Clinical Chemistry.