Explicit measurement of the endotoxin adsorption efficiency detects non-Langmuir behavior at low concentrations

Ekaterina G Kholina; Alexey M Nesterenko; Tatiana V Galochkina; Danil A Gvozdev; Irina V Polyakova; Ilya B Kovalenko; Marina G Strakhovskaya; Oleg A Pisarev

doi:10.1016/j.ab.2019.113445

Explicit measurement of the endotoxin adsorption efficiency detects non-Langmuir behavior at low concentrations

Anal Biochem. 2019 Dec 15:587:113445. doi: 10.1016/j.ab.2019.113445. Epub 2019 Sep 19.

Authors

Ekaterina G Kholina¹, Alexey M Nesterenko², Tatiana V Galochkina³, Danil A Gvozdev¹, Irina V Polyakova⁴, Ilya B Kovalenko⁵, Marina G Strakhovskaya⁶, Oleg A Pisarev⁷

Affiliations

¹ M.V.Lomonosov Moscow State University, Biology Faculty, Leninskie Gory 1/12, Moscow, 119234, Russia.
² A.N.Belozersky Institute of Physico-Chemical Biology, MSU, Leninskie Gory 1/40, Moscow, 119991, Russia; M.V.Shemyakin and Yu.A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Science, Miklukho-Maklaya str. 16/10, Moscow, 117997, Russia. Electronic address: comconadin@gmail.com.
³ Université de Paris, UMR_S1134, BIGR, Inserm, Institut National de Transfusion Sanguine, DSIMB, 6 rue Alexandre Cabanel, 75739, Paris, France.
⁴ Institute of Macromolecular Compounds, Russian Academy of Science, Bolshoi pr. 31, Saint-Petersburg, 199004, Russia.
⁵ M.V.Lomonosov Moscow State University, Biology Faculty, Leninskie Gory 1/12, Moscow, 119234, Russia; Federal Research and Clinical Center of Specialized Medical Care and Medical Technologies, Federal Medical and Biological Agency of Russia, Orekhovy bld. 28, Moscow, 115682, Russia; Astrakhan State University, Tatischeva str. 20a, Astrakhan, 414056, Russia; Scientific and Technological Center of Unique Instrumentation of the Russian Academy of Sciences, Butlerova str. 15, Moscow, 117342, Russia.
⁶ M.V.Lomonosov Moscow State University, Biology Faculty, Leninskie Gory 1/12, Moscow, 119234, Russia; Federal Research and Clinical Center of Specialized Medical Care and Medical Technologies, Federal Medical and Biological Agency of Russia, Orekhovy bld. 28, Moscow, 115682, Russia.
⁷ Institute of Macromolecular Compounds, Russian Academy of Science, Bolshoi pr. 31, Saint-Petersburg, 199004, Russia; Peter the Great St. Petersburg Polytechnic University, 195251, Polytechnicheskaya 29, St. Petersburg, Russia.

PMID: 31542342
DOI: 10.1016/j.ab.2019.113445

Abstract

Lipopolysaccharides (LPS) are the Gram-negative bacteria cell wall components capable to induce the system inflammatory response even at picomolar concentrations. LPS detection at these concentrations is necessary to develop new sorbents for the efficient purification of the biological fluids. LAL-test widely used for LPS concentration estimation is based on the LPS biological activity measurement and thus may depend on the LPS concentration in a non-linear way. Here we propose a new explicit method for the LPS concentration measurement based on fluorescently labeled LPS and direct photon counting and develop the new protocol for LPS adsorption efficiency measurement. Following the suggested protocol in the experiments on novel sorbents, we demonstrate that LPS adsorption at small biologically relevant concentrations is non-Langmuir.

Keywords: Adsorption; Direct photon counting; Endotoxin; Fluoresence; Lipopolysaccharides; Sepsis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adsorption
Lipopolysaccharides / analysis*
Molecular Structure

Substances

Lipopolysaccharides