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2002 02 [Abstract][OnlineView][Download 27k] - ZHANG Aihua, BI Lan, WANG Zhiyou et al( Wuhan Institute of Biological Products, Wuhan 430060)
[Abstract] Objective To obtain the VH and VL variable region genes of murine McAbs to human CD molecule. Methods The VH and VL variable region genes were amplified from the total RNA of WuT3, WuT4, WuTac, WuLCA and WuT9 hyridoma cells by RT - PCR separately, then cloned and sequenced. Results All the length of the PCR products were about 400bp. Restriction enzyme analysis proved that cloning vectors pMDIS - T - VL and pMDIS - T - VH were successfully constructed, and the sequencing results of them were in accord with expected. Conclusion All the obtained fragments were proved to be ahtibody genes in comparison with Kabat and Blast antibody gene homology database.
2002 02 [Abstract][OnlineView][Download 685k] - BAI Yang , ZHANG Yali, WANG Jide et al( Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071)
[Abstract] Objective To obtain the heat shock protein 60(Hsp60)gene of Helicobacter pylori and clone it into plasmid pET - 22b ( + ) for nucleotide sequencing. Methods Hsp60 gene was amplified by PCR,then inserted into pET- 22b( + ) vector and sequenced.Results The sequence of Hsp60 gene was in agreement with that reported in Genebank. Conclusion A confirmed Hsp60 gene has been obtained in this paper.The study laid a good foundation of the recombination,expression and research of the gene.
2002 02 [Abstract][OnlineView][Download 569k] - JIANG Wenzheng , JIN Ningyi, WANG Hongwei et al ( Key Laboratory of Gene Engineering, Quartermaster University of PIA , Changchun 130062)
[Abstract] Objective To construct a recombinant plasmid pPICGAG for the expression of whole length of HIV - 1 Gag gene in Pichia pastoris . Methods The plasmid pKSGAG containing the whole length of HIV - 1 Gag gene was digested with Not I and Xhol and cloned into expression vector pPIC9. The constructed plasmid pPICGAG was linarized with Sac I and transformed to Pichia pastoris GS 115.The positive transforma-nts were identified by PCR and expressed in BMGY and BMMY media.The expressed products were analyzed by SDS - PAGE. Results The integration rate of transformant was 72.7% . SDS - PAGE showed a relative molecular weight of expressed protein of about 55000, and Western blot showed specific reaction of it with McAb. Conclusion The HIV - 1 Gag protein with good reactogenicity and specificity was successfully expressed in Pichia pastoris.
2002 02 [Abstract][OnlineView][Download 1678k] - LIU Yongqing , CHEN Puyan, DU Nianxing et al( Key Laboratory of Animal Diseases Diagnosis and Immunity , Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095)
[Abstract] Objective To construct E.coli K99 pilus expression vector.Methods E. coli K99 pi-lus forming subunit gene(fanC) was cloned by a series of subcloning method and sequenced. The mutation site was determined by the epitope mimesis of fanC subunit. Results A gene fragment containing fanC and a part of fanD genes was cloned. The fragment was significantly longer than that reported abroad. Sequencing showed a new gene fragment at the binding region of fanC and fanD. Compared with the E. coli gene in Genebank, the new fragment was identified as the inserted sequence IS1 with a length of 768bp. No obvious change was observed in the reading frame of fanD gene after IS1 was inserted. However,8 amino acids were added at the end of fanC fragment. The epitope mimesis of fanC subunit showed an obvious hydrophilic region in K99 fane C fragment. According to the previous experiences, the region was judged as the site in which epitope was located. Conclusion Inserted sequence IS1 was observed in E. coli K99 gene for the first time.
2002 02 [Abstract][OnlineView][Download 1013k] - TONG Kuitang , JIN Mei, QIAN Chunhong et al ( Shanghai Institute of Biological Products, Shanghai 200052)
[Abstract] Objective To study on the molecular structure and biological activity of recombinant human IFN-7. Methods The biological activity of IFN-7 manufactured by Shanghai Institute of Biological Products (SIBP) was detected by cytopathy inhibition and calculated into international unit using Japanese IFN-y reference as standard, and the relative molecular weight of it was detected by 4 methods. Results The specific activity of the IFN-γmanufactured by SIBP was not less than 3.0 ×107 IU/mg protein,and the relative molecular weight of it was lower than 16900. It consisted of only 129 amino acids,and the sequences of 15 to 19 amino acids at the N - teminal of it were in accord with those in theory. Conclusion The deletion of amino acids was observed at the C - tenninal of IFN-y manufactured by SEBP. However, the biological activity of the EFN-γhas not been influenced.
2002 02 [Abstract][OnlineView][Download 142k] - WU Jie,XU Gelin, ZHENG Xinxiong et al( Wuhan Insitute of Biological Products, Wuhan 430060)
[Abstract] Objective To study on the immunological characteristics of 4 rabies street virus strains isolated in China. Methods The antigenicities and immunogenicities of the 4 strains were detected by immunity and neutralizing tests. Results Significant difference were observed in the antigeinicities of the 4 strains. However, the vaccine prepared with aG strain could protect mice against the challenge with the 4 street virus strains. Conclusion aG vaccine showed good cross immunity to the 4 street virus strains.
2002 02 [Abstract][OnlineView][Download 118k] - ZHANG Li, JIN Yihui, ZHANG Yuanxing( State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237)
[Abstract] Objective To observe the influence of by-product( ammonia) in serum - free medium on the growth and metabolism of hybridoma cells. Methods Add the ammonia at various concentrations and observe the growth and metabolism of hybridoma cells and the production of McAbs. Results The growth and metabolism of hybridoma cells were influenced after only 0.8 mmol/L of ammonia was added in. The ammonia at a concentration of 4 mmol/L showed significant inhibiting effect on the growth and metabolism of hybridoma cells. When the concentration reached above 6 mmoI/L,the growth was stopped, and the metabolism was also influenced seriously. Along with the addition of ammonia, the concentration of McAb in culture supernatant was decreased,however, the specific production rate of it was increased. Conclusion Ammonia showed significant influence on the growth and metabolism of hybridoma cells, and the influence of it on McAb concentration was mainly caused by the decrease of cell density.
2002 02 [Abstract][OnlineView][Download 236k] - HAN Liang, WANG Qi, RAN Shu et al ( Changchun Institute of Biological Products , Changchun 130062)
[Abstract] Objective To establish a simple method suitable for the large - scale purification of inactivated hemorrhagic fever with renal syndrome (HFRS) vaccine. Methods The inactivated HFRS vaccine prepared with primary golden hamster kidney cells (GHKC) was purified by various methods such as ultrafiltra-tion, precipitation with zinc acetate, affinity, ion - exchange and gel filtration chromatography, and the results of them were compared. Results The purities of vaccines purified ultrafiltration plus Sepharose 4FF column chromatography or precipitation widi zinc acetate plus Sepharose 4FF column chromatography met Chinese Requirements for Biologies. However, those purified by ceEufine sulfate gel affinity or ion- exchange chromatography were unsatisfied. Conclusion An economic and simple method suitable for the large - scale purification of inactivated HFRS vaccine is established.
2002 02 [Abstract][OnlineView][Download 222k] - LIN Gang,XU Shijia, YANG Bangling et al( Wuhan Institute of Biological Products, Wuhan 430060)
[Abstract] Objective To develop a novel material and a new method for preparing bovine nerve growth factor. Methods The nerve growth factor was purified from bovine seminal plasma by a two - step cation exchange chromatography. Results The relative molecular weight, specific activity and electrophoretic purity of the purified bovine nerve growth factor were 30000,400000BU/rag and above 95% respectively, and a single peak was observed on the FPLC profile of it. Conclusion Nerve growth factor can be purified from bovine seminal plasma.
2002 02 [Abstract][OnlineView][Download 655k] - LIU Hong , PAN Hongchun et al ( Southwest Pharmaceutical Company of Limited Liability, Chongqing 400038)
[Abstract] Objective To establish a simple and effective procedure for the separation and purification of L - asparaginase from E. coli and study on the characteristics of purified product. Methods L - aspargin-ase was separated with dilute alkaline liquid from E. coli broken with cold acetone,purified by ion exchange and affinity chromatography and characterized by various methods. Results The pruity, specific activity and total recovery rate of L- asparaginase were 94.4% ,520u/mg and 65.7% respectively.Both the L- asparaginase with relative molecular weights of 143900 and 166700 were EC - 2 components with anti - tumor activities and showed the maximum absorptions at a wavelength of 270nm. The optimal temperature and pH value of it were40to50℃ and 8.0 to 9.0 respectively.The L-asparaginase was stable at 40℃l below.Conclusion An appropriate procedure for the separation and purification of L - asparaginase from E. coli was established.
2002 02 [Abstract][OnlineView][Download 823k]
2002 02 [Abstract][OnlineView][Download 77k] - BAI Jianshi , BU Fengrong, LI Song et al( National Institute for the Control of Pharmaceutical and Biological Products, Beijing 100050)
[Abstract] Objective To explore the impossibility of preparing erythrocyte substiute by the selective modification of bovine hemoglobin(bHb) with mPEG. Methods Bovine hemoglobin was modified with mPEG, and the influence of mPEG on the physicochemical property of it was studied. Results The mean coupling rate of mPEG and bHb reached 30% ,and the eletrophoretic migration rate, absorption spectrum and colloid osmotic pressure of bHb were significantly changed. Conclusion The preparation of erythrocyte substitute by modifying bHb with mPEG is worth of further study.
2002 02 [Abstract][OnlineView][Download 1595k]
2002 02 [Abstract][OnlineView][Download 80k] - LIU Chuanxuan , MA Qingjun, ZHANG Yanhong et al( Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100850)
[Abstract] Objective To prepare oral subunit B/whole cell vaccine for preventing cholera caused by strain 0139.Methods The subunit B/ whole cell vaccine was prepared with the subunit B of recombinant cholera toxin and the 0139 strains of vibrio cholerae inactivated with formalin,and the safety and immunogenic-ity of it were studied. Results No bodyweight loss, morbidity or death were observed in the mice immunized with the oral vaccine at large dosages for 3 days. All the mice were survival and healthy. The specific antibody of high liters were detectecd out in the peripheral blood and intestinal tract of mice immunized with the vaccine by oral route or I. P. injection. The vaccine protected the mice against the challenge with live 0139 strain. Conclusion The subunit B/whole cell vaccine showed good safety and immunogenicity in mice.
2002 02 [Abstract][OnlineView][Download 978k]
2002 02 [Abstract][OnlineView][Download 63k] - HOU Jifeng, CHENG Yaqin (National Institute for the Control of Pharmaceutical and Biological Products, Beijing 100050)
[Abstract] Objective To detect the biological activity of human serum high density lipoprotein correctly. Methods The physiological function of HDL stimulating cholesterol (Ch) efflux from cells were detected by U937 foaming cell model. Protein and cholesterol contents were detected by Brad Ford method and Ch kit respectively, and the amounts of Ch stimulated with HDL at different dosages were compared. Results The method showed good relativity (r≥.98) within certain limits of HDL concentration. The reproducibility and recovery of it were 6.1% and 91.8%-110.5% respectively. Conclusion The method can reflect the function of HDL transferring cholesterol from cells more directly. Using human - derived cells, the method is simple to handle and easy to be spread.
2002 02 [Abstract][OnlineView][Download 135k]
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2002 02 [Abstract][OnlineView][Download 75k] - CHEN Hongping, PAN Huiming, HE Huiqing et al ( Yichang Sanitation and Anti - epidemic Station, Yichang 443000)
[Abstract] Objective To control the infections and morbidities of hepatitis A and B and improve the immunity of population of the next generation. Methods A practical management mode of planned immunization with hepatitis A and B vaccines was developed according to the epidemic and immunological characteristics of hepatitis A and B as well as the immune state of population in Yichang City, Hubei Province. The management mode including a series of scientific and systemic requirements for vaccination subject, immune schedule, dosage, vaccination route, transport, supply and statistical data of hepatitis A and B vaccines were implemented by the planned immunization net of the whole city. Results After the management mode was implemented , vaccination cards and certificates were distributed to 99.52% and 98.34% of the children in the city respectively, and the immunization rates of hepatitis A and B vaccines reached 96.25% and 98.30% respectively. Compared with those before implement, the morbidity of hepatitis A and B decreased by 92.63% (from 289.687 100,000 to 21.35/ 100,000) and 64.60% (from 200.37/100,000 to 70.94/100,000) respectively, and the serum anti - HAV and anti - HBV positive rates increased from 33.80% and 42.72% to 91.57% and 88.76% respectively. No outbreak of hepatitis A or B occurred during 1998 - 2000 in the places in which children gathered, such as kindergartens and schools. Conclusion The implement of the management mode has gained good social benefit and promoted the development of planned immunization and disease control.
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2002 02 [Abstract][OnlineView][Download 363k] 下载本期数据