A test-system based on enzyme-linked immunosorbent assay (ELISA) for quantitative determination of cyclosporin A (CSA) in human whole blood has been developed. The detection limit of the method was 25 ng/ml, the linearity of the method in the concentration range of 60–1400 ng/ml varied from 94 to 105%, the variation coefficient did not exceed 8%. The novel method exhibited good correlation with radioimmunoassay and polarization fluoroimmunoassay methods; the linear regression coefficients were 0.965 and 0.984, respectively. The developed test system is stable for at least 9 months when stored at 4°C and can be used in clinical practice.

The methodical bases for detecting antibiotics using a bioluminescent assay and blood serum are briefed. Antibiotics inhibit the luminescence of a genetically engineered Escherichia coli strain. The degree of inhibition depended on the type of antibiotic, its concentration, and the time of cell incubation with antibiotic. The highest cell sensitivity was recorded towards the aminoglycoside antibiotics, which amounted to 85 ± 10 ng/ml for gentamicin and streptomycin. The sensitivity of this system to a number of antibiotics essentially increased when the cells were previously activated with blood serum. The sensitivity of this method for gentamicin and streptomycin in the presence of blood serum amounted to 2.5 ± 0.5; for tetracycline, 45 ± 8 ng/ml. Use of the sera containing specific antibodies to the antibiotic detected provided a high sensitivity of the biosensor tested. Comparison of the luminescences of E. coli cells activated with normal and specific antisera upon incubation with an antibiotic allows the type of antibiotic and its quantitative content in the sample to be determined. Characteristic of the analysis of antibiotics with the help of recombinant E. coli are a high accuracy, sensitivity, specificity, simplicity, and a short time needed for measurement.

The conjugation of the drugs with vector molecules enables to obtain therapeutic preparation, which may be transported to the selected target organ. In the present work the methods of conjugation of antineoplastic enzyme L-lysine alpha-oxidase with antibodies were elaborated. Conjugates were worked out through the attachment of amino groups on the antibody surface either with the aldehyde groups which were created in L-lysine alpha-oxidase molecule (0.2% of initial enzymatic activity) or with the aldehyde groups of cross-linking molecules. Maximal (78%) L-lysine alpha-oxidase activity in conjugates was observed when oxidized peroxidase which contained the aldehyde groups was used as crosslinking agent. The glutaraldehyde method yielded 70% of initial enzyme activity.

Оглавление: 1. Ферменты 1.1. Щелочная фосфатаза 1.2. Аминотрансферазы 1.3. α-Амилаза 1.4. Лактатдегидрогеназа 1.5. Креатинкиназа 1.6. γ-Глутамилтрансфераза 2. Субстраты и белки 2.1. Мочевина 2.2. Креатинин 2.3. Мочевая кислота 2.4. Белок общий 2.5. Альбумин 2.6. Билирубин 2.7. Глюкоза 2.8. Холестерин и фракции холестерина 2.9. Триглицериды 2.10. Железо, ЛЖСС, ОЖСС 2.11. Кальций 2.12. Магний 2.13. Калий 2.14. Натрий 2.15. Хлориды 2.16. Фосфор неорганический 2.17. Гликозилированный гемоглобин 2.18. С-реактивный белок 3. Правила подготовки пациента для проведения биохимических исследований 4. Литература

The coupled peroxidase-catalysed chemiluminescence reaction of luminol and p-iodophenol oxidation of H2O2 (the 'enhanced-chemiluminescence reaction') was presence of synthetic polyelectrolytes with various chemical addition of both aliphatic poly-(NN'-dimethyldiallylammonium chloride) (PDM) polycation and aromatic poly-(N-ethyl-4-vinylpyridinium bromide) polycation (PEVP) in the reaction mixture led to an increase in the kinetic constants for the elementary enzymic reaction of luminol oxidation. By contrast, whereas the addition of PDM gave rise to an increase in chemiluminescence light intensity (I-CL), the addition of PEVP caused a decrease in the signal. The results of a study of a model non-enzymic reaction proved that the observed decrease of ICL is due to the interaction of PEVP pyridinium groups with the intermediate products of luminol oxidation. The most pronounced decrease of ICL was in the case of the polyelectrolyte complex formed by PEVP polycation and the conjugate of peroxidase with poly(methacrylic acid) (polyanion). The demonstrated ability of synthetic polyelectrolytes to effect both enzymic and non-enzymic stages of the reaction could be used in the development of new immunoassays of biologically active substances.