A protein biosensor for lactate.
Blood lactate is a clinically valuable diagnostic indicator. In this preliminary report we describe a protein biosensor for L-lactate based on beef heart lactate dehydrogenase (LDH). LDH was noncovalently labeled with 8-anilino-1-naphthalene sulfonic acid (ANS). The ANS-labeled LDH displayed an approximately 40% decrease in emission intensity upon binding lactate. This decrease can be used to measure the lactate concentration. The ANS-labeled LDH was further utilized in a new sensing format, polarization sensing, which is suitable for miniaturization to a point-of-care lactate monitor. However, temporal instability of beef heart LDH indicates the need for further protein engineering prior to development of a more robust lactate-sensing protein.
D'Auria
S
S
Department of Biochemistry and Molecular Biology, Center for Fluorescence Spectroscopy, University of Maryland at Baltimore, 21201, USA.
Gryczynski
Z
Z
Gryczynski
I
I
Rossi
M
M
Lakowicz
J R
JR
D000817:Anilino Naphthalenesulfonates / Q000378:metabolism; D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods*; D002417:Cattle; D004305:Dose-Response Relationship, Drug; D005456:Fluorescent Dyes / Q000378:metabolism; D007527:Isoenzymes; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D019344:Lactic Acid / Q000097:blood*; D015233:Models, Statistical; D009206:Myocardium / Q000201:enzymology; D013050:Spectrometry, Fluorescence
D000817:Anilino Naphthalenesulfonates; D005456:Fluorescent Dyes; D007527:Isoenzymes; D019344:Lactic Acid; C027132:1-anilino-8-naphthalenesulfonate; D007770:L-Lactate Dehydrogenase