Biocatalytically initiated precipitation atom transfer radical polymerization (ATRP) as a quantitative method for hemoglobin detection in biological fluids

Jonas Pollard, Omar Rifaie-Graham, Samuel Raccio, Annabelle Davey, Sandor Balog, Nico Bruns

Research output: Contribution to journalArticle

Abstract

The hemoglobin content of blood is an important health indicator, and the presence of microscopic amounts of hemoglobin in places where it normally does not occur, e.g. in blood plasma or in urine, is a sign of diseases such as hemolytic anemia or urinary tract infections. Thus, methods to detect and quantify hemoglobin are important for clinical laboratories, blood banks, and for point-of-care diagnostics. The precipitation polymerization of N-isopropylacrylamide by hemoglobin-catalyzed atom transfer radical polymerization (ATRP) is used as an assay for hemoglobin quantification relying on the formation of turbidity as a simple optical read-out. Dose-response curves for pure hemoglobin and for hemoglobin in blood plasma, in urine, in erythrocytes, and in full blood are obtained. Turbidity formation increases with the concentration of hemoglobin. Concentrations of hemoglobin as low as 6.45 × 10-3 mg mL-1 in solution, 4.88 × 10-1 mg mL-1 in plasma, and 1.65 × 10-1 mg mL-1 in urine could be detected, which is below the clinically relevant concentrations in the respective body fluids. Total hemoglobin in full blood is also accurately determined. The reaction can be regarded as a polymerization-based signal amplification for the sensing of hemoglobin, as the analyte catalyzes the formation of radicals which add many monomer units into detectable polymer chains. While most established hemoglobin tests involve the use of highly toxic reagents such as potassium cyanide, the polymerization-based test uses simple and stable organic reagents. Thus, it is an environmentally friendlier alternative to established chemical assays for hemoglobin.

Original languageEnglish
Pages (from-to)1162-1170
Number of pages9
JournalAnalytical Chemistry
Volume92
Issue number1
Early online date2 Dec 2019
DOIs
Publication statusE-pub ahead of print - 2 Dec 2019

Fingerprint

Atom transfer radical polymerization
Hemoglobins
Fluids
Blood
Polymerization
Turbidity
Plasmas
Assays
Potassium Cyanide
Clinical laboratories
Poisons
Body fluids
Amplification
Polymers
Monomers

Keywords

  • hemoglobin
  • ATRP
  • biocatalytic ATRP
  • polymerization-based amplification
  • anemia diagnostic
  • biosensing
  • precipitation polymerization

Cite this

Pollard, Jonas ; Rifaie-Graham, Omar ; Raccio, Samuel ; Davey, Annabelle ; Balog, Sandor ; Bruns, Nico. / Biocatalytically initiated precipitation atom transfer radical polymerization (ATRP) as a quantitative method for hemoglobin detection in biological fluids. In: Analytical Chemistry. 2019 ; Vol. 92, No. 1. pp. 1162-1170.
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Biocatalytically initiated precipitation atom transfer radical polymerization (ATRP) as a quantitative method for hemoglobin detection in biological fluids. / Pollard, Jonas; Rifaie-Graham, Omar; Raccio, Samuel; Davey, Annabelle; Balog, Sandor; Bruns, Nico.

In: Analytical Chemistry, Vol. 92, No. 1, 02.12.2019, p. 1162-1170.

Research output: Contribution to journalArticle

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T1 - Biocatalytically initiated precipitation atom transfer radical polymerization (ATRP) as a quantitative method for hemoglobin detection in biological fluids

AU - Pollard, Jonas

AU - Rifaie-Graham, Omar

AU - Raccio, Samuel

AU - Davey, Annabelle

AU - Balog, Sandor

AU - Bruns, Nico

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N2 - The hemoglobin content of blood is an important health indicator, and the presence of microscopic amounts of hemoglobin in places where it normally does not occur, e.g. in blood plasma or in urine, is a sign of diseases such as hemolytic anemia or urinary tract infections. Thus, methods to detect and quantify hemoglobin are important for clinical laboratories, blood banks, and for point-of-care diagnostics. The precipitation polymerization of N-isopropylacrylamide by hemoglobin-catalyzed atom transfer radical polymerization (ATRP) is used as an assay for hemoglobin quantification relying on the formation of turbidity as a simple optical read-out. Dose-response curves for pure hemoglobin and for hemoglobin in blood plasma, in urine, in erythrocytes, and in full blood are obtained. Turbidity formation increases with the concentration of hemoglobin. Concentrations of hemoglobin as low as 6.45 × 10-3 mg mL-1 in solution, 4.88 × 10-1 mg mL-1 in plasma, and 1.65 × 10-1 mg mL-1 in urine could be detected, which is below the clinically relevant concentrations in the respective body fluids. Total hemoglobin in full blood is also accurately determined. The reaction can be regarded as a polymerization-based signal amplification for the sensing of hemoglobin, as the analyte catalyzes the formation of radicals which add many monomer units into detectable polymer chains. While most established hemoglobin tests involve the use of highly toxic reagents such as potassium cyanide, the polymerization-based test uses simple and stable organic reagents. Thus, it is an environmentally friendlier alternative to established chemical assays for hemoglobin.

AB - The hemoglobin content of blood is an important health indicator, and the presence of microscopic amounts of hemoglobin in places where it normally does not occur, e.g. in blood plasma or in urine, is a sign of diseases such as hemolytic anemia or urinary tract infections. Thus, methods to detect and quantify hemoglobin are important for clinical laboratories, blood banks, and for point-of-care diagnostics. The precipitation polymerization of N-isopropylacrylamide by hemoglobin-catalyzed atom transfer radical polymerization (ATRP) is used as an assay for hemoglobin quantification relying on the formation of turbidity as a simple optical read-out. Dose-response curves for pure hemoglobin and for hemoglobin in blood plasma, in urine, in erythrocytes, and in full blood are obtained. Turbidity formation increases with the concentration of hemoglobin. Concentrations of hemoglobin as low as 6.45 × 10-3 mg mL-1 in solution, 4.88 × 10-1 mg mL-1 in plasma, and 1.65 × 10-1 mg mL-1 in urine could be detected, which is below the clinically relevant concentrations in the respective body fluids. Total hemoglobin in full blood is also accurately determined. The reaction can be regarded as a polymerization-based signal amplification for the sensing of hemoglobin, as the analyte catalyzes the formation of radicals which add many monomer units into detectable polymer chains. While most established hemoglobin tests involve the use of highly toxic reagents such as potassium cyanide, the polymerization-based test uses simple and stable organic reagents. Thus, it is an environmentally friendlier alternative to established chemical assays for hemoglobin.

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