Abstract

Conjugates of gold nanoparticles (GNPs) and antibodies are widely used in various fields of biochemistry and microbiology. However, the procedure for obtaining such conjugates remains precarious, and the properties of conjugates differ significantly for different antibody clones. One of the most common problems is the aggregation of GNPs in the course of their conjugation with antibodies. This article considers an example of the conjugation of monoclonal antibodies with non-stable aggregating product. The composition of the antibody preparation was studied using electrophoresis, asymmetrical flow field-flow fractionation, and ultracentrifugation. It was shown that the component that causes the aggregation of the GNPs is the light chains of immunoglobulins that appear due to the spontaneous decay of the antibodies. After separation of the fraction with a molecular weight of less than 30 kDa, stable conjugates of antibodies with GNPs were obtained. The high functional activity of the obtained conjugates was confirmed by immunochromatography.

Highlights

  • Due to the unique physical and chemical properties of gold nanoparticles (GNPs), they are widely used in biochemistry, medicine, and analytical chemistry [1,2,3,4,5,6]

  • Dimensional characteristics of GNPs were evaluated by transmission electron microscopy

  • Dimensional characteristics of GNPs were evaluated had by transmission microscopy

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Summary

Introduction

Due to the unique physical and chemical properties of gold nanoparticles (GNPs), they are widely used in biochemistry, medicine, and analytical chemistry [1,2,3,4,5,6]. GNPs have high surface energy, which determines the possibility of their modification [7,8]. GNPs are modified using biological macromolecules such as proteins and nucleic acids [9,10]. A large number of covalent sewing techniques have been developed, but for biological macromolecules, physical adsorption methods remain the most popular [13,14,15]. The interaction of macromolecules and nanoparticles depends on the chemical nature of the adsorbed substance, the composition of the medium, and other factors.

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