RESUMEN
A number of drugs based on recombinant erythropoietin contain human serum albumin as an auxiliary component. The presence of this protein hinders the proper control of the drug quality in accordance with the requirements of regulating agencies. We propose the novel method for separation of recombinant erythropoietin (epoetin beta) and human serum albumin. It is based on the subsequent use of hydrophobic sorbent and anion exchange resin placed in gravity flow columns (without the use of spin-columns). The proposed approach makes it possible to concentrate and purify the preparations containing the epoetin beta both at high and at minimal concentrations (the ratio of the amount of albumin and erythropoietin in the used preparations can reach 125:1). The average yield of epoetin beta after the use of hydrophobic sorbent and anion exchange resin was 75% and 97%, respectively. It was shown that the determined conditions of sample preparation had no affect on the content of the epoetin beta in the product.
RESUMEN
AIMS: To demonstrate the effectiveness of application of the adsorbent successively modified with nano-layers of fluoroplast and polyaniline for one-step isolation of DNA of hepatitis B virus and transfusion-transmitted virus from human serum. MATERIALS & METHODS: The technique is based on the application of the spin-cartridges containing porous adsorbent for one-step viral DNA isolation from serum followed by polymerase chain reaction. RESULTS: The developed adsorbent was shown to be effective for one-step isolation of viral DNA from serum samples for polymerase chain reaction diagnostics. CONCLUSION: The effectiveness of the developed adsorbent application for isolation of viral DNA from serum for polymerase chain reaction diagnostics was confirmed in comparison with standard methods. Thus, the facile sample preparation method of viral DNA isolation was elaborated.
Asunto(s)
ADN Viral/análisis , Virus de la Hepatitis B/genética , Reacción en Cadena de la Polimerasa/métodos , Adsorción , Patógenos Transmitidos por la Sangre , HumanosRESUMEN
AIMS: A new approach for the preparation of adsorbents for one-step isolation/purification of DNA from different samples (e.g., bacterial lysates, smears and blood) has been developed. MATERIALS & METHODS: The technique is based on the use of porous silica preliminary treated with ozone followed by grafting of polymer nanolayers on the activated carrier surface. The chemical nature of active centers, their stability and conditions for the use of the activated support as heterogeneous initiator of different macromolecular reactions were studied. RESULTS: The adsorbents modified with thin (~3.0-7.5 nm) polytetrafluoroethylene and polyaniline layers were prepared and characterized. Sorption properties of the obtained adsorbents were demonstrated on examples of express (2-5 min) one-step DNA isolation for direct use in PCR diagnostics. CONCLUSION: The effectiveness of the developed adsorbents used for DNA isolation and purification was confirmed in comparison with the standard methods. Thus, a facile (alternative in relation to irradiation postpolymerization or standard oxidative polymerization techniques) manufacturing method of the materials for nucleic acids sample preparation was developed.