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OBJECTIVE: To evaluate the effect of health education on schistosomiasis in Kunshan City, so as to provide the evidence for making the consolidating strategy in the late stage of interruption of schistosomiasis transmission. METHODS: The residents, middle school students and elementary school students were randomly sampled from one community, one middle school and one elementary school of each of two towns and they were investigated with interviews and questionnaires for the implementation of health education on schistosomiasis prevention and control. RESULTS: A total of 452 middle school students (232 cases) and primary school students (220 cases) were surveyed. The awareness rate of total schistosomiasis knowledge was 98.21% among the students (the awareness rate of basic schistosomiasis knowledge was 98.42% and the awareness rate of preventive schistosomiasis knowledge was 98.01%). Among the 220 elementary school students, the awareness rate of total schistosomiasis knowledge was 97.21% (the awareness rate of basic schistosomiasis knowledge was 97.60% and the awareness rate of preventive schistosomiasis knowledge was 96.82%). Among the 232 middle school students, the awareness rate of total schistosomiasis knowledge was 99.17% (χ2 = 34.661, compared with the rate of the elementary school students) [the awareness rate of basic schistosomiasis knowledge was 99.20% (χ2 = 13.045, compared with the rate of the elementary school students) and the awareness rate of preventive schistosomiasis knowledge was 99.14% (χ2 = 21.796, compared with the rate of the elementary school students)]. There were significant differences between the elementary school students and middle school students in above-mentioned awareness rates (all P < 0.001). There were schistosomiasis health education materials or teaching plans in all the four schools. Among the 402 residents surveyed, the awareness rate of total schistosomiasis knowledge was 98.87%. CONCLUSIONS: The effect of health education on schistosomiasis prevention and control is very well, and the total awareness rate of schistosomiasis prevention and control knowledge among the population has reached the goal (more than 95%) of the medium- and long-term planning of schistosomiasis prevention and control in Kunshan City.
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Educación en Salud , Conocimientos, Actitudes y Práctica en Salud , Esquistosomiasis/prevención & control , China , Humanos , Instituciones Académicas , Estudiantes , Encuestas y CuestionariosRESUMEN
OBJECTIVE: To establish the quality control system for the recombinant human GLP-1 analogue fusion protein. METHODS: The potency of the fusion protein was determined by luciferase reporter gene assay. The purity was analyzed by non-reduced SDS-PAGE and SEC-HPLC respectively. RP-HPLC was used for the peptide mapping. ELISA was used to analyze the identification of the final products. The molecular mass and peptide mass spectra were analyzed by LC-ESI-MS technique. Other control tests were performed according to the requirements in the Chinese Pharmacopoeia (Volume III, 2010 edition). RESULTS: Control tests were performed on three different lots of bulks and final products of recombinant GLP-1 analog fusion protein by the developed methods. The results showed that all the indexes met the requirements in the Guideline for Quality Control of Recombinant DNA Products for Human Use and Chinese Pharmacopoeia (Volume III, 2010 edition). The molecular weight of the recombinant human GLP-1 analogue fusion protein was 71 361.0, which was in conformity with the theoretical value. CONCLUSION: The developed methods and standard may assure the safety, effectiveness, and controllability of the recombinant human GLP-1 analogue fusion protein, which might be used for the routine quality control of products of the same kind.
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The study aims to characterize and compare interferon reference standards from 5 manufacturers. By testing molecular mass and trypsin-digested peptide mass mapping, the amino acid sequence was verified and post-translational modifications such as disulfide bond were identified. Results show that the molecular mass and amino acid sequence were consistent with theory; the disulfide bonds of 4 lots of interferon were Cys1-Cys98/Cys29-Cys138, 1 lot was Cys29-Cys139/Cys86-Cys99; N-terminal "+Met", acetyl N-terminal and Met oxidation were identified in part of the sample. UPLC-MS can be used to characterize and compare interferon reference standards from different manufacturers.
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Secuencia de Aminoácidos , Cromatografía Líquida de Alta Presión , Métodos , Interferones , Estándares de Referencia , Espectrometría de Masas , Métodos , Peso Molecular , Oxidación-Reducción , Mapeo Peptídico , Procesamiento Proteico-Postraduccional , Estándares de ReferenciaRESUMEN
To establish a detection method of oncolytic adenovirus/p53 and standard of quality control, human telomerase reverse transcriptase (hTERT) promoter, CMV fusion promoter containing hypoxia reaction element (HRE) and p53 gene were identified by vector DNA restriction enzyme digestion and PCR analysis. The result conformed that all modified regions were in consistent with theoretical ones. Particle number was 2.0 x 10(11) mL(-1) determined by UV (A260). Infectious titer was 5.0 x 10(10) IU mL(-1) analyzed by TCID50. In vitro p53 gene expression in human lung cancer cell H1299 was determined by ELISA, and A450 ratio of nucleoprotein in virus infection group to control group was 5.2. Antitumor potency was evaluated by cytotoxicity assay using human lung cancer cell A549, and the MOI(IC50) of this gene therapy preparation was 1.0. The tumor cells targeted replication ability of recombinant virus was determined by TCID50 titer ratio of filial generation virus between human lung cancer cell A549 and human diploid epidermal fibrolast BJ cells after infected by virus with same MOI. TCID50 titer ratio of tumor cell infection group to normal cell infection control group was 398. The IE-HPLC purity of virus was 99.5%. There was less than 1 copy of wild type adenovirus within 1 x 10(7) VP recombinant virus. Other quality control items were complied with corresponding requirements in the guidance for human somatic cell therapy and gene therapy and Chinese pharmacopeia volume III. The detection method of oncolytic adenovirus/p53 was successfully established for quality control standard. The study also provided reference for quality control of other oncolytic viral vector products.
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Humanos , Adenoviridae , Genética , Metabolismo , Fisiología , Línea Celular Tumoral , Regulación Neoplásica de la Expresión Génica , Genes p53 , Terapia Genética , Vectores Genéticos , Neoplasias , Metabolismo , Patología , Virología , Virus Oncolíticos , Genética , Metabolismo , Fisiología , Control de Calidad , Proteínas Recombinantes de Fusión , Genética , Metabolismo , Transfección , Replicación ViralRESUMEN
To establish methods and requirements for quality control of rhLFA3-IgG1, biological potency of rhLFA3-IgG1 was determined by CD2 molecule competitive binding assay on Jurkat cell surface. Purity of rhLFA3-IgG1 was analyzed by SEC-HPLC and IEC-HPLC. Peptide mapping was preformed by tryptic digestion and RP-HPLC after sample reduced and carboxymethylation by DTT and indoacetic acid, respectively. CHO host cell protein and Protein A residual were detected by ELISA separately. The quality control methods and requirements, such as biological potency, the physical-chemical characteristic of rhLFA3-IgG1 had been established. The methods and requirements for quality control of rhLFA3-IgG1 showed advantages of assuring the products safety and efficacy, which can be used for routine quality control of rhLFA3-IgG1.
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Humanos , Unión Competitiva , Biotecnología , Métodos , Antígenos CD2 , Metabolismo , Antígenos CD58 , Química , Cromatografía Líquida de Alta Presión , Inmunoglobulina G , Química , Células Jurkat , Peso Molecular , Mapeo Peptídico , Control de Calidad , Proteínas Recombinantes de Fusión , QuímicaRESUMEN
<p><b>AIM</b>To analyze the peptide mapping of recombinant human interleukin-11 (rhIL-11) by HPLC-ESI-Q-TOF/MS spectrometry.</p><p><b>METHODS</b>The trypsin digested rhIL-11 at 37 degrees C over night, and the peptide mapping was performed by HPLC. The relative molecular weight of the peptides fragments was measured by ESI-Q-TOF/MS, and amino acid sequence was analyzed by MS/MS.</p><p><b>RESULTS</b>The peptide fragments of rhIL-11 in the peptide mapping were assigned by analyzing the retain time, relative molecular weight and amino acid sequence. And 97% of the expected peptides were detected in this way.</p><p><b>CONCLUSION</b>The study proves that HPLC-ESI-Q-TOF/MS is a good method to analyze peptide mapping of protein with the advantage of sensitivity, high speed and accuracy.</p>
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Secuencia de Aminoácidos , Cromatografía Líquida de Alta Presión , Métodos , Interleucina-11 , Química , Genética , Peso Molecular , Fragmentos de Péptidos , Mapeo Peptídico , Métodos , Proteínas Recombinantes , Química , Reproducibilidad de los Resultados , Espectrometría de Masa por Ionización de Electrospray , Métodos , Espectrometría de Masas en Tándem , MétodosRESUMEN
<p><b>AIM</b>To establish methods and requirements for quality control of recombinant human tumor necrosis factor receptor Fc fusion protein (rhTNFR-Fc).</p><p><b>METHODS</b>Biological potency of rhTNFR-Fc was determined by neutralizing the bioactivity of TNF-alpha. rhTNFR-Fc samples were reduced by beta-mercaptoethanol and the peptide map was performed by tryptic digestion. Residual protein A and the host cell protein content were detected by ELISA. Anti-TNFR and anti-Fc antibodies were used in ELISA for detection of the rhTNFR-Fc content.</p><p><b>RESULTS</b>The quality control methods, such as bioassay, peptide map, residual protein A detection, were established and used for quality control of rhTNFR-Fc. The unit of rhTNFR-Fc (AU) was defined according to the international unit of TNF-alpha. The specific activity was up to 8 x 10(4) AU.mg-1. The requirements for quality control of rhTNFR-Fc were established.</p><p><b>CONCLUSION</b>The methods and requirement were used for quality control of rhTNFR-Fc products.</p>
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Animales , Ratones , Biotecnología , Métodos , División Celular , Etanercept , Inmunoglobulina G , Química , Farmacología , Mapeo Peptídico , Control de Calidad , Receptores del Factor de Necrosis Tumoral , Química , Proteínas Recombinantes de Fusión , Química , Farmacología , Tecnología Farmacéutica , Estándares de Referencia , Células Tumorales CultivadasRESUMEN
<p><b>AIM</b>To establish the quality control methods for recombinant human endostatin.</p><p><b>METHODS</b>Biological activity was determined by endothelial cell migration assays. Peptide mapping was tested by trypsin digestion and RP-HPLC. Purity was determined by non-reduced SDS-PAGE and RP-HPLC. Other tests including molecular weight, isoelectrical point, etc. were done according to the National Requirements for Biological Products (2000).</p><p><b>RESULTS</b>The method of bioassay was established and used for determining activity of endostatin. Specific activity of the three batchs of drug substance was 1.45 x 10(6), 1.57 x 10(6) and 2.73 x 10(6) u.mg-1 proteins. Peptide mappings of the three batches of drug substance were completely identified. Both purity results of the products tested by SDS-PAGE and RP-HPLC were more than 99%.</p><p><b>CONCLUSION</b>The established methods can effectively control the quality of recombinant human endostatin.</p>