Measurement of biosynthesis and breakdown rates of biological molecules that are inaccessible or not easily accessible to direct sampling, non-invasively, by label incorporation into metabolic derivatives and catabolitic products
AssigneeThe Regents of the University of California
InventorMarc K. Hellerstein
Life of this patent
Estimating the term…
Abstract
Methods of determining rate of biosynthesis or breakdown of biological molecules from metabolic derivatives and catabolic products are disclosed herein. In particular, methods of measuring the rates of biosynthesis and breakdown of biological molecules inaccessible or not easily accessible to direct sampling by sampling metabolic derivatives and catabolic products in accessible biological samples are disclosed herein.
Claims59
A method for determining the rate of biosynthesis or breakdown of one or more inaccessible biological molecules in an individual comprising the steps of:
- a) administering an isotope-labeled precursor molecule to an individual for a period of time sufficient for the label of said isotope-labeled precursor molecule to become incorporated into said one or more inaccessible biological molecules;
- b) obtaining one or more accessible biological samples from an individual, wherein said one or more accessible biological samples comprise one or more metabolic derivatives of said one or more inaccessible biological molecules as resulting from in vivo metabolism of said inaccessible biological molecules;
- and
- c) detecting the incorporation of said label in said one or more metabolic derivatives by mass spectrometry to determine said rate of biosynthesis or breakdown of said one or more inaccessible biological molecules.
A kit for determining the biosynthetic rate or breakdown rate of one or more inaccessible biological molecules in an individual comprising:
- a) an isotope-labeled precursor, and
- b) instructions for use of the kit,
- wherein the kit is used to determine the biosynthetic rate or breakdown rate of said one or more inaccessible biological molecules in said individual.
A method of measuring the rate of biosynthesis or breakdown of a biological molecule comprising:
- administering an isotope-labeled precursor molecule to an individual;
- obtaining an accessible biological sample comprising one or more metabolic derivatives of said inaccessible biological molecule from said individual;
- measuring the proportion of labeled to unlabeled metabolic derivatives at one or more times;
- calculating the isotope enrichment of said inaccessible biological molecule by mass isotopomer distribution analysis (MIDA);
- and
- applying precursor-product or exponential decay equations to determine said rate of biosynthesis or breakdown of said inaccessible biological molecule.
A method for determining the rate of biosynthesis or breakdown of collagen in an individual comprising the steps of:
- a) administering deuterated water to an individual for a period of time sufficient for said deuterium to become incorporated into said collagen;
- b) obtaining one or more accessible biological samples from an individual, wherein said one or more accessible biological samples comprise one or more collagen derivatives selecting from the group consisting of pyridinoline, deoxypyridinoline, hydroxyproline, hydroxylysine, glucosylgalactosyl-hydroxylysine, galactosylhydroxylysine, N-terminal telopeptide α(I) (SEQ ID NO:
- 1), N-terminal telopeptide α2(I) (SEQ ID NO:
- 2), N-terminal telopeptide α2(I) (SEQ ID NO:
- 3), N-terminal telopeptide α1(II) (SEQ ID NO:
- 4), N-terminal telopeptide α1(III) (SEQ ID NO:
- 5), C-terminal telopeptide α1(I) (SEQ ID NO:
- 6), C-terminal telopeptide α2(I) (SEQ ID NO:
- 7), C-terminal telopeptide α1(II) (SEQ ID NO:
- 8), C-terminal telopeptide α1(II) (SEQ ID NO:
- 9), C-terminal telopeptide α1(II) (SEQ ID NO:
- 10), C-terminal telopeptide α1(III) (SEQ ID NO:
- 11), cross-linked carboxy-terminal peptide of type I collagen (ICTP), PINP(α1) (SEQ ID NO:
- 12), PICP(α1) (SEQ ID NO:
- 13), PINP(α2) (SEQ ID NO:
- 14), PICP(α2) (SEQ ID NO:
- 15), PIINP(α1) (SEQ ID NO:
- 16), PIICP(α1) (SEQ ID NO:
- 17), PIIINP(α1) (SEQ ID NO:
- 18), PIIICP(α1)(SEQ ID NO:
- 19), PIVNP(α1)(SEQ ID NO:
- 20), PIVNP(α2)(SEQ ID NO:
- 21), PIVNP(α2)(SEQ ID NO:
- 22), PIVNP(α3) (SEQ ID NO:
- 23), PIVNP(α4) (SEQ ID NO:
- 24), PIVNP(α5) (SEQ ID NO:
- 25), and PIVNP(α6) (SEQ ID NO:
- 26);
- c) detecting said one or more collagen derivatives by mass spectrometry;
- d) calculating the isotope enrichment of said one or more collagen derivatives by mass isotopomer distribution analysis (MIDA);
- and
- e) applying precursor-product or exponential decay equations to determine the rate of biosynthesis or breakdown of said collagen.
Dependencies are inferred from the claim text.