Open AccessNew immunoassay systems based on recombinant human lactoferrin
Author(s) -
Denis A. Semenov,
И. И. Вашкевич,
О. В. Свиридов
Publication year - 2021
Publication title -
doklady nacionalʹnoj akademii nauk belarusi
Language(s) - English
Resource type - Journals
eISSN - 2524-2431
pISSN - 1561-8323
DOI - 10.29235/1561-8323-2021-65-3-290-302
Subject(s) - lactoferrin , chemistry , horseradish peroxidase , immunoassay , recombinant dna , conjugate , antiserum , affinity chromatography , chromatography , biochemistry , enzyme , antibody , biology , mathematical analysis , mathematics , gene , immunology
In this work, soluble and solid phase immunoreagents, including recombinant human lactoferrin (rhLF), a complex of rhLF with europium ions, rabbit antiserum to rhLF, anti-rhLF immunoglobulin purified by antigen-affinity chromatography and the conjugates of this immunoglobulin with an Eu3+ chelate or horseradish peroxidase have been obtained by a combination of biochemical and synthetic methods using rhLF as an initial compound. Biospecific interactions of the reagents in four immunochemical systems were assessed by measuring the enzyme activity or time-resolved fluorescence. The study resulted in the development of fast and precise immunoassays for biologically active rhLF in transgenic goat milk and in protein fractions obtained in the course of pure rhLF manufacture, as well as in pharmaceutical preparations and food additives.