RESUMO
A method was developed for the rapid detection of human epidermal growth factor based on a sandwich-format immunochromatographic assay. The contact between the sample and the test strip with immobilized immunoreagents initiates the fluid flow movement across the membrane components of the test strip, immunochemical reactions, and the formation of colored lines. Requirements on the configuration of the test system in order to achieve the lowest limit of detection were defined in the course of the development of the assay. It was shown that this method enables the detection of human epidermal growth factor within 5 min at concentrations as low as 10 pg/mL in aqueous solutions, urine, and the blood serum and plasma. The developed test system can be used for point-of-care diagnostics.
Assuntos
Anticorpos Monoclonais Murinos/química , Cromatografia de Afinidade/métodos , Fator de Crescimento Epidérmico/sangue , Fator de Crescimento Epidérmico/urina , Membranas Artificiais , Cromatografia de Afinidade/instrumentação , Humanos , Sensibilidade e EspecificidadeRESUMO
The approaches for high sensitive and specific determination of nonylphenol (NP) with the help of immune enzymatic (ELISA) method have been developed. The process of preparation of conjugates of NP with proteins, antiserum obtaining, purification of immunoglobulin (Ig) fractions and study of specificity of the obtained antibodies were described in detail. It was shown that the antiserum and total Ig fraction do not differ in respect of specificity and binding abilities. The developed algorithm of ELISA method fulfillment is able to provide NP revealing with the sensitivity in the range of 20-50 ng/ml and working controlled concentrations--from 20-50 to 1000 ng/ml.
