# US 2008/0187937 A1 · Method For Measuring Dynamics Of Self-Assembling Systems Of Biological Molecules In Vivo And Uses For Discovering Or Evaluating Therapeutic Agents > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2008/0187937 A1 - Document ID: US20080187937A1 - Status: Published application - Kind: A1 - Application number: 11/573,374 - Filed: 2005-08-08 - Published: 2008-08-07 - Earliest claimed: 2004-08-07 - Inventors: Marc K. Hellerstein; Patrizia Fanara - Web page: https://patentgrep.com/patent/US20080187937A1 ## CPC - G01N 33/58: involving labelled substances (G01N33/53 takes precedence) - G01N 33/6896: {Neurological disorders, e.g. Alzheimer's disease} - G01N 2458/15: Non-radioactive isotope labels, e.g. for detection by mass spectrometry - G01N 2500/00: Screening for compounds of potential therapeutic value - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) ## Abstract The Applicants have established a simple, rapid assay of measuring the dynamics of self-assembling systems of biological molecules, based on stable isotope labeling technology that can be used in intact animals including humans. Examples of self-assembling systems of biological molecules include microtubule polymers, actin filaments, amyloid-beta plaques or fibrils, prion plaques or fibrils, fibrin aggregates, tau filaments (e.g., neurofibrillary tangles), α-synuclein filaments, and mutant hemoglo-bin aggregates. The method reveals constitutive differences in the dynamics of assembly and disassembly between tissues and is sensitive to the action of compounds that stabilize these dynamics. ## Claims ### Claim 1 A method for measuring the rate of self assembly of subunits into biological molecular assemblages in a test living system as compared to a control living system, said method comprising: a) administering an isotope-labeled substrate to said living system for a first period of time sufficient for said substrate to be incorporated into at least one of said subunits and at least one of said molecular assemblages; b) obtaining a first sample from said living system; c) quantifying the amount of labeled molecular assemblages from said first sample; d) quantifying the amount of unincorporated labeled subunits; e) comparing the amount of labeled molecular assemblages to the amount of labeled molecular assemblage in a control living system; and f) comparing the amount of unincorporated labeled subunits to the amount of unincorporated labeled subunits in a control living system; to determine a difference in said rate of self assembly in said test living system as compared to said control living system. ### Claim 2 (depends on 1) The method according to claim 1 further comprising calculating the molecular flux rate of said labeled molecular assemblages, wherein said comparing step comprises calculating the ratio of said rates and comparing said ratio to the ratio of molecular flux rates in said control living system. ### Claim 3 (depends on 1) The method according to claim 1 wherein said isotope-labeled substrates are tubulin protein dimers. ### Claim 4 (depends on 1) The method of claim 1, wherein said isotope-labeled subunits are monomeric actin proteins. ### Claim 5 (depends on 1) The method of claim 1, wherein said isotope-labeled subunits are prion proteins. ### Claim 6 (depends on 1) The method of claim 1, wherein said isotope-labeled subunits are amyloid-beta proteins. ### Claim 7 (depends on 1) The method of claim 1, wherein said isotope-labeled subunits are fibrin proteins. ### Claim 8 (depends on 1) The method of claim 1, wherein said isotope-labeled subunits are mutant hemoglobin proteins. ### Claim 9 (depends on 1) A method according to claim 1 further comprising administering a candidate agent to said test living system. ### Claim 10 (depends on 9) A method according to claim 9 wherein said candidate agent is administered prior to said administration of said isotope-labeled substrate. ### Claim 11 (depends on 9) A method according to claim 9 wherein said candidate agent is administered during said administration of said isotope-labeled substrate. ### Claim 12 (depends on 9) A method according to claim 9 wherein said candidate agent is administered after said administration of said isotope-labeled substrate. ### Claim 13 (depends on 1) A method according to claim 1 further comprising administering said substrate for a second period of time and repeating steps b)-f). ### Claim 14 (depends on 1) A method according to claim 1 further comprising obtaining a second sample and repeating steps c)-f). ### Claim 15 (depends on 5) A methods according to claim 5 wherein said candidate agent is a drug. ### Claim 16 (depends on 1) The method of claim 1, wherein said isotope-labeled substrate is labeled with a stable isotope. ### Claim 17 (depends on 1) The method of claim 1, wherein said isotope-labeled substrate is stable-isotope labeled water. ### Claim 18 (depends on 17) The method of claim 17, wherein said stable-isotope labeled water is labeled with 2H. ### Claim 19 (depends on 1) The method of claim 1, wherein said isotope-labeled substrate is labeled with a radioactive isotope. ### Claim 20 (depends on 19) The method of claim 19, wherein said isotope-labeled substrate is radioactive-isotope labeled water. ### Claim 21 (depends on 20) The method of claim 20, wherein said radioactive isotope-labeled water is labeled with 3H. ### Claim 22 (depends on 1) The method of claim 1, wherein said test living system and said control living system are mammals. ### Claim 23 (depends on 1) The method of claim 1, wherein said test living system and said control living system are human. ### Claim 24 (depends on 1) The method of claim 1, wherein said test living system and said control living system are cell lines. ### Claim 25 (depends on 1) The method of claim 1, wherein said test living system and said control living system are primary cells. ### Claim 26 (depends on 25) The method of claim 25, wherein said primary cells are sperm cells. ### Claim 27 (depends on 1) The method of claim 1, wherein said test living system and said control living system are bacterial cells.