Using size-exclusion chromatography to quantify the 146S antigen in inactivated foot-and-mouth disease vaccine / 生物工程学报

The aim of this study is to quantify the 146S antigen in foot-and-mouth disease virus (FMDV) inactivated vaccine by size-exclusion chromatography (SEC). The analysis was performed on a TSKgel G4000SWXL column (7.8 mm×30 cm), with a pH 7.2 buffer salt system as the mobile phase. The flow rate was 0.6 mL/min, the injection volume was 100 μL and the detection wavelength was 259 nm. The calibration curve was established by using purified inactivated FMDV (serotype O) 146S antigen; 3 batches of vaccine formulated by inactivated antigen solution were tested to verify the accuracy, reproducibility, specificity and tolerability of the method. At last 16 batches of vaccine were determined by the SEC method. Results showed a good linearity between peak area and concentration of 146S antigen in the range between 0.56 and 67.42 μg/mL (R2=0.996, n=10), and the average recovery rate of 146S antigen in the 3 batches of vaccine formulated in lab were 93.6% (RSD=2.7%, n=3), 102.3% (RSD=2.6%, n=3), and 95.5% (RSD=5.1%, n=3). The method was proved accurate and reliable with good reproducibility (RSD=0.5%, n=6), and applied to determine 16 batches of the commercial FMDV vaccine. According to the above results, the SEC method is high effective for 146S antigen quantify in the inactivated FMDV vaccine and would provide strong support for the vaccine quality control.

Cloning and Sequencing of NorB Gene of Nitrobacterium / 环境与健康杂志

Objective To amplify and sequence the distinctive norB gene of Nitrobacterium. Methods The norB gene was amplified by PCR and was cloned into T-vector and then transformed into E.coli JM109. After white-and-blue selection, positive colonies were sequenced with T7 sequencing primers by Takara Company. Spliced with Vector NTI Suite software, the homologous analysis of sequences were conducted in Genbank through Internet. Results The results showed that all of norB genes obtained from the nitrite-oxidizing bacteria were homologous with the norB gene of Nitrobacterium hamburgensis in Genbank. The homology ranged from 96% to 98%, and there were no frameshift mutations, which guaranteed the correct ORF. Conclusion The obtained genes are norB genes indeed.

Preliminary Screen and Identification of One Nitrobacteria / 环境与健康杂志

Objective To screen and seperate nitrobacteria that could be used to treat nitrogenous compound pollution.Methods Autotrophic nitrite-oxidizing microorganisms were screened by enrichment culture from soil.After morphological and biological detection,primers were designed to detect the distinctive gene norB of nitrobacteria.The PCR production was se-quenced,and the homology was identified according to the sequence in Genbank.Results One strain of bacterium was screened that could oxidize nitrite.This bacterium was small,light-yellow bacilli.The expected-size DNA fragments could be amplicated by PCR.The sequence of a PCR production was blasted in Genbank and showed99%consistent with norB gene of N hamburgensis.Conclusion It was preliminarily confirmed that the microorganism screened in this study might be nitrobac-terium.

Screening and Preliminary Identification of Nitrifying Bacteria in Soil / 环境与健康杂志

ve To control and minimize the water pollution by nitrogen. Methods The autotrophic amino-oxidizing bacteria in soil were screened by enrichment and incubation. After bacterio-morphological identifica-tion their nitrifying rates were determined. Results All of the 12 screened strains were gram-negative and in the shape of brevi-rod dorminantly. The colonies with a dorminant form of short diameter in round shape, light yellow and smooth formed after 7-day incubation at 28℃ . The nitrifring rates ranged 0.40 mg/ (L?d)~1. 15 mg/ (L?d) . Conclusion The screened 12 strains all presented nitrifying function, some of them could be used to control water environment pollution by nitrogen.