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Biomedical subjects

A M Egorov

Publications and source records attributed to A M Egorov.

At least 37 records · Page 2Linked to original sources
PubMed2000

Oriented immobilization of antibodies onto the gold surfaces via their native thiol groups.

The general approach for site-oriented immobilization of antibodies onto gold supports is reported. The immobilization is carried out using the native sulfide groups of immunoglobulin (IgG). To liberate the thiol groups, the intact IgG was split into two half-IgG fragments without destruction of the binding site of the antibody. The immobilization of half-IgG fragments on the gold surface was carried out by simple adsorption. The antigen binding capacity of the half-IgG modified gold supports is similar to that of the gold surfaces with the traditionally linked antibodies and is much higher than for nonspecifically adsorbed intact IgGs. The immobilized antibodies, according to the proposed approach, maintain high antigen binding constants. The immobilization procedure provides orientation of IgG fragments in terms of the similar distance between the binding site of the antibody and the surface of the gold support, which does not cause the distribution of the apparent affinity constants. The high operational stability of half-IgG modified gold electrodes makes them applicable for analytical applications.

Electrochemistry↗
PubMed1999

Chemiluminescent multiassay of pesticides with horseradish peroxidase as a label.

Competitive chemiluminescent immunoassay based on a combination of five antibodies was used in a combination with neural network to identify and estimate amounts of three cross-reacting s-triazines (atrazine, terbythylazine and ametryn). Antibodies with different cross-reactivity towards s-triazines were immobilized in separate wells an eight-well microtiter strip. Training of neural networks was carried out with four different learning procedures. The best topology for the data measured was a net with two hidden layers with ten neurons in the first and 15 in the second layer trained with the Schmidhuber method. s-Triazine classification of environmental samples containing various analyte mixtures was correct in 70-100% of all cases depending on the type of analyte. The test developed can be proposed as an alternative field test for multianalyte environmental monitoring.

Atrazine↗
PubMed1999

[Optimization of a method of solid-phase immunoenzyme analysis for determination of chloramphenicol in milk].

The method of solid-phase enzyme linked immunosorbent assay (ELISA) for quantitative detection of chloramphenicol (CAP) in milk was developed. Peculiarities of the adsorption on the microtitre plates surface of CAP-ovalbumin conjugate were investigated. Different conditions of competition stage of the analysis were studied. Conditions providing CAP monitoring in human blood serum in the clinical range were optimized. Matrix effect on the assay results was studied. The specificity of the analytical system was investigated and the reagents stability was examined. The method developed permits CAP concentration to be determined in human blood serum, diluted 1/100, in the linear range from 0.1 to 100 ng/ml. The assay is characterized by high sensitivity (0.08 ng/ml) and good reproducibility (CV < 10.8%), assay time is about 3 hours. The correlation coefficient with HPLC is 0.977.

Animals↗
PubMed1998

Chemiluminescent biosensors based on porous supports with immobilized peroxidase.

Combining a sensitive and fast chemiluminescence detection method with novel immobilization methods for horseradish peroxidase (HRP) has enabled us to develop sensitive sensors for hydrogen peroxide and the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D). HRP biospecific immobilization was shown to have well-defined advantages in terms of a higher specific activity of the enzyme of the surface and a greater sensitivity in the enhanced chemiluminescence (ECL) reaction. The lower detection limit for hydrogen peroxide was 0.025 nmol with HRP immobilized on nitrocellulose membrane through avidin-biotin linkage. An immunosensor for the detection of 2,4-D provides a lower detection limit of 0.2 microgram/l with specific antibodies covalently immobilized on photoactivated nylon. The assay with chemiluminescent detection was also characterized with a wider concentration range when compared with the colorimetric assay.

2,4-Dichlorophenoxyacetic Acid↗
PubMed1998

[Development of a solid-phase immunoenzyme analysis of gentamicin in human blood serum].

An enzyme immune test system was designed and optimized for quantitative assay of gentamicin in human sera. Immunospecific reagents i.e. gentamicin conjugates with ovalbumin (for sorption on polysterol plates) and with bovine serum albumin (immunogen) were prepared. Gentamicin specific antisera were isolated and tested. Conditions for the antigen sorption on polysterol plates were determined and optimized. Different regimes of the competition reaction were investigated and conditions for the antibiotic assay in human sera were determined. The assay specificity was studied and the stability of the test system was checked. An experimental lot of the reagent set was manufactured at the ZAO NPP Immunotech and the correlation tests with the use of the fluorescence polarization immunoassay were performed. The set is destined for the assay of 40 samples (in duplicate). The method sensitivity is 1 ng/ml of gentamicin. The range of the detectable concentration is 1 to 32 ng/ml of gentamicin in 1000-fold diluted sera. The assay time is not more than 3 hours. The variation coefficient of the results does not exceed 12 per cent. The shelf-life of the set is 0.5 years when stored at a temperature of 2 to 8 degrees C.

Anti-Bacterial Agents↗
PubMed1997

Epitope mapping of horseradish peroxidase (isoenzyme C).

Peptide scanning (PEPSCAN) was used to determine linear antigenic determinants of horseradish peroxidase isoenzyme C (HRPC). For this purpose, we synthesized 303 overlapping hexapeptide fragments (with a step of one amino acid residue) of the protein primary structure and studied their interactions with anti-HRPC polyclonal antisera by ELISA. Experiments with various titers of antisera allowed us to determine linear antigenic determinants of HRPC; several such determinants were spatially located in regular elements of the secondary structure (alpha-helices) found both inside and outside the protein globule. A fraction of epitopes were located in loops and folds of the HRPC peptide chain with irregular shapes. These epitopes contained several functionally important residues: Arg 38, which is part of the active site of the enzyme, as well as Phe 142 and Phe 143, which form a channel allowing aromatic substrates to reach the active site. Amino acid residues that form calcium-binding sites or occur in the vicinity of disulfide bonds are not involved in these epitopes.

Amino Acid Sequence↗