WO2001077140A3 - Measurement of nutrient uptake in cells and methods based thereon - Google Patents

Measurement of nutrient uptake in cells and methods based thereon Download PDF

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Publication number
WO2001077140A3
WO2001077140A3 PCT/US2000/028913 US0028913W WO0177140A3 WO 2001077140 A3 WO2001077140 A3 WO 2001077140A3 US 0028913 W US0028913 W US 0028913W WO 0177140 A3 WO0177140 A3 WO 0177140A3
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WO
WIPO (PCT)
Prior art keywords
glucose
cell
uptake
rates
sugar
Prior art date
Application number
PCT/US2000/028913
Other languages
French (fr)
Other versions
WO2001077140A2 (en
Inventor
Srienc Friederich
Arvind Natarajan
Nicholas R Abu-Absi
Original Assignee
Srienc Friederich
Arvind Natarajan
Abu Absi Nicholas R
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Srienc Friederich, Arvind Natarajan, Abu Absi Nicholas R filed Critical Srienc Friederich
Priority to AU59015/01A priority Critical patent/AU5901501A/en
Publication of WO2001077140A2 publication Critical patent/WO2001077140A2/en
Publication of WO2001077140A3 publication Critical patent/WO2001077140A3/en

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/5308—Immunoassay; Biospecific binding assay; Materials therefor for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites

Abstract

The fluorescent glucose analog, 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose (2-NBDG), was used to measure rates of glucose uptake by single Escherichia coli cells. When cell populations were exposed to the glucose analog, the sugar was actively transported and accumulated in single cells to a steady-state level that depended upon the extracellular concentration of the sugar, the sugar transport capacity of the cells, and the intracellular degradation rate. The dependence upon substrate concentration could be described according to Michaelis-Menten kinetics with apparent saturation constant KM=1.75 νM, and maximum uptake rate = 197 molecules/cell-second. Specificity of glucose transporters to the analog was confirmed by inhibition of uptake of 2-NBDG by D-glucose, 3-o-methyl glucose, and D-glucosamine, and lack of inhibition by L-glucose. The assay for sugar uptake is extremely sensitive such that the presence of even trace amounts of D-glucose in the culture medium (∩0.2 νM) is detectable. The rates of single-cell sugar uptake were found to increase differentially with cell size as measured by microscopy or single-cell light scattering intensity. A mathematical model was developed to provide a theoretical basis for estimating single-cell glucose uptake rates from single-cell 2-NBDG uptake rates. Because the distribution of single-cell sugar uptake rates of the entire cell population is measured, this assay provides a novel means of estimating the instantaneous rates of nutrient depletion in the culture medium.
PCT/US2000/028913 1999-10-19 2000-10-19 Measurement of nutrient uptake in cells and methods based thereon WO2001077140A2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU59015/01A AU5901501A (en) 1999-10-19 2000-10-19 Measurement of nutrient uptake in cells and methods based thereon

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US16033599P 1999-10-19 1999-10-19
US60/160,335 1999-10-19

Publications (2)

Publication Number Publication Date
WO2001077140A2 WO2001077140A2 (en) 2001-10-18
WO2001077140A3 true WO2001077140A3 (en) 2002-02-21

Family

ID=22576473

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2000/028913 WO2001077140A2 (en) 1999-10-19 2000-10-19 Measurement of nutrient uptake in cells and methods based thereon

Country Status (2)

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AU (1) AU5901501A (en)
WO (1) WO2001077140A2 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111073897B (en) * 2020-01-14 2021-04-27 华中农业大学 Application of GhTMT2 gene in regulation of soluble sugar accumulation in cotton

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ES2302605B2 (en) * 2006-03-02 2009-05-01 Universidade De Santiago De Compostela METHOD FOR THE DETECTION OF GLUCOGENO BY FLUORESCENCE.
JP6013735B2 (en) 2008-09-30 2016-10-25 マリンクロット ニュークリア メディシン エルエルシー Conjugates of hexoses and metal coordination bonds for imaging purposes
BR112012009828B8 (en) 2009-10-26 2022-10-25 Hoffmann La Roche METHOD FOR THE PRODUCTION OF A GLYCOSYLED IMMUNOGLOBULIN AND ITS USE
EP3246709A3 (en) 2010-09-16 2018-02-14 The General Hospital Corporation Red blood cell dynamics for diagnosis
WO2017106461A1 (en) * 2015-12-15 2017-06-22 The General Hospital Corporation Methods of estimating blood glucose and related systems
US11293852B2 (en) 2016-04-07 2022-04-05 The General Hospital Corporation White blood cell population dynamics

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5643883A (en) * 1995-01-19 1997-07-01 Uab Research Foundation Glucose-6-phosphate uptake inhibitors and novel uses thereof
US6207136B1 (en) * 1997-03-11 2001-03-27 Tokyo University Of Agriculture And Technology Fluorescent imaging method of saccharide uptake activity of living tissue

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5643883A (en) * 1995-01-19 1997-07-01 Uab Research Foundation Glucose-6-phosphate uptake inhibitors and novel uses thereof
US6207136B1 (en) * 1997-03-11 2001-03-27 Tokyo University Of Agriculture And Technology Fluorescent imaging method of saccharide uptake activity of living tissue

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111073897B (en) * 2020-01-14 2021-04-27 华中农业大学 Application of GhTMT2 gene in regulation of soluble sugar accumulation in cotton

Also Published As

Publication number Publication date
WO2001077140A2 (en) 2001-10-18
AU5901501A (en) 2001-10-23

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