- Zhang Shumin *,Hu Benzhong,Zhuang Hui et al( * National Institute for the Control of Pharmaceutical and Biological Products,Beijing 100050 )
Objective To explore the roles of different HCV antigens in detection and the relationship between them with HCV RNA.Methods The serum samples from HCV carrier and the patients with acute,chronic hepatitis and HCV related hepatocellular carinoma were detected for the antibodies to antigens encoded by the different gene fragments of HCV and HCV RNA by EIA and RT PCR respectively.Results Compared with those of anti E1,E2,NS4 and NS5,the positive rates of anti NS3 and anti C were the highest in the 4 groups of patients.A proportion of the patients in each group was anti NS4 and/or anti NS5 positive.The positive rate of total anti HCV was much higher than that of antibody to the antigen encoded by any gene fragment of HCV.Conclusion NS3 and C antigens play important roles in the detection of anti HCV.However,the role of NS4 and NS5 antigens in EIA should not be omitted.Combined use of different HCV will improve the sensitivity of EIA significantly.
2000 04 [Abstract][OnlineView][Download 30k] - Ma Li *,Wang Xiaoning et al( * Institute of Molecular Immunology,The First Military Medical University,Guangzhou 510515 )
Objective To express human vascular endothelial growth factor receptor FLT 1 extracellular domain 1 3 loop cDNA in Pichia.pastoris ,purify the expressed product and detect the biological activity of it .Methods By inserting human FLT 1(1 3 loop) cDNA coding 316 amino acid residues into Pichia.pastoris expression vector pPIC9K containing AOXI promoter and the sequences of α secreting signal peptides,a recombinant expression plasmid pPIC9K/FLT 1(1 3) was constructed and transformed to GS115,then His +Mut s phenotype transformant was selected and cultured in flasks,and FLT 1(1 3)was expressed under the induction of 1% methanol.Results SDS PAGE showed that,after being induced with 1% methanol for 4 days, the expressed product existed in supernatant in the form of soluble molecule and contained 60% of total protein expressed.Western blot showed good antigenicity and specificity of expressed product.After being purified by CM Sepharose FF and Sephacryl S 100 chromatography,the purity of expressed product reached above 90%.Biological assay proved that the expressed product could combine with hVEGF165 and inhibit the proliferation of HUVEC stimulated by hVEGF 165.Conclusion Human vascular endothelial growth factor receptor FLT 1 extracellular domain (1 3 loop)was successfully expressed.The study laid a foundation of further application of the expressed product in the treatment of vasoformationrelated diseases,such as tumor and diabetic retinopathy.
2000 04 [Abstract][OnlineView][Download 88k] - Zeng Xianwu,Hu Xiaofang,Wang Qianchuan et al( Cheng du Institute of Biological Products,Chengdu 610063 )
Objective For highly expression of human 125 Ser IL 2.Methods Human 125 Ser IL 2 gene was amplified by PCR,then cloned into transposition vector and expressed using a novel Bac to Bac baculovirus expression system.The immunological and biological activities of expressed protein were detected by Western blot and CTLL 2/MTT method respectively.Results The expressed protein showed the immunological and biological activities of human IL 2.SDS PAGE proved that the expression level of the protein reached 23.47% .Conclusion Human 125 Ser IL 2 is highly expressed in insect cells.
2000 04 [Abstract][OnlineView][Download 60k] - Mao Jianping,Dong Yan,Liu Bin et al( Beijing Institute of Radiation Medicine,Beijing 100850 )
Objective To prepare anti GPA McAb for establishing GPA somatic mutation assay.Methods The McAb to human GPA N was prepared with the spleen cells of BALB/c mice immunized with O, N type human erythrocytes and purified GPA N by hybridoma technique, selected by cell agglutination and indirect immuno fluorescence microscopy, and identified by ELISA, Western blot and indirect immuno fluorescence with enzyme treated human erythrocytes.Results A McAb cell strain 6A8(IgG3,λ) recognizing the epitopes at sites 1 8 of GPA N was obtained. The strain was dependent on glycosyl chain. Flow cytometric selection showed that 6A8 specifically bound to MN and N type human erythrocytes fixed with dimethyleneimine.Conclusion The study has theoretical and practical values in analyzing the GPA N mutation caused by gene mutation and glycosylation variation.
2000 04 [Abstract][OnlineView][Download 77k] - Jia Lili *,Yu Yongxin,Theodere F.Tsai et al( * National Institute for the Control of Pharmaceutical and Biological Products,beijing 100050 )
Objective To study on the immunity of live attenuated Japanese Encephalitis(JE)vaccine to different wild JE virus strains.Methods The neutralizing effect of the vaccine on different wild JE virus strains was detected by plaque reduction neutralization test(PRNT),and the immunogenicity of it was studied in mice by vaccination challenge protection test.Results The vaccine could protect mice against the challenge with 11 JE virus strains isolated in China and 11 in Thailand,Vietnam,Indonesia,India,Philippines and Japan.The protective rate of it reached above 90%.Human immune sera showed significant neutralizing effect on 1 JE virus strain isolated in Taiwan and 9 strains isolated abroad.Conclusion Live attenuated JE vaccine prepared with strain SA14 14 2 has broad spectrum immunogenicity.
2000 04 [Abstract][OnlineView][Download 33k] - Zhu Wei,Liu Weiqing,Ma Yingjie( Shanghai Institute of Biological Products,Shanghai 200052 )
Objective To explore if the purified Jiangzhe agkistrodon halys antivenom prepared by the authors could neutralize Korean snake venoms.Methods The cross immunological reaction of agkistrodon halys antivenom and Korean snake venoms were detected by double immuno diffusion test,and the neutralizing capacity of each milliliter of antivenom to both the lethal and hemorrhagic toxicity of 3 kinds of Korean snake venoms were calculated by statistical method according to the results of animal test.Results Each milliliter of antivenom could neutralize 1 0 3.9 and 2.5mg of A.saxntitis,A.brevicauclus and A.usserience lethal venom respectively;it can also neutralize 0.8,3.15 and 2.0mg of haemorrhagic venom of the 3 kinds of snakes respectively.Conclusion Jiangzhe agkistrodon halys antivenom can neutralize Korean snake venoms.
2000 04 [Abstract][OnlineView][Download 48k] -
2000 04 [Abstract][OnlineView][Download 10k] - Wang Renli, Yang Yiming, Zhong Ping( Shanghai Institute of Biological Products, Shanghai 200052)
Objective To prepare a kit for detecting apoprotein(Apo) A1 and B. Methods Dextran sulfate precipitation, ultracentrifugation, chromatography, and so on. Results Either Apo A1 and B separated by the above-mentioned methods was proved to be a single component. Anti-Apo Al and anti-Apo B sera were obtained by immunizing sheep with Apo Al and B respectively. The detection results of the 2 kinds of sera were consistent with those of imported products(Bochinger, Germany), and the kit for turbidimetry prepared with them showed good correlation with those manufactured by Diasys Co.(Germany). Conclusion The kit reached the quality level of imported product of the same kind.
2000 04 [Abstract][OnlineView][Download 51k] - Wei Jingyan * et al( * Jilin Province Center for Clinical Loboratory, Changchun 130021)
Objective To prepare the McAb to cardiac troponin Ⅰ(cTnⅠ) for further preparation of diagnostic kit for acute myocardiac infarction(AMI).Methods TnC-Sepharose 4B affinity chromatography column was prepared with rabbit skeletal muscles troponin C(sInC) and used for the direct purification of cTn Ⅰ from human cardiac muscle tissue.Results SDS-PAGE proved that the cTnⅠ was electrophoretic pure, and the molecular weight and amino acid component of it were consistent with that reported in documents.Conclusion The study lays a foundation of developing the diagnostic kit for early AMI.
2000 04 [Abstract][OnlineView][Download 37k] - Chen Weijin *,Sun Hong,Guan Feng et al( *Changchun Institute of Biological Products,Changchun 130062 )
Objective To prepare Lys plasminogen and study on the stability of it.Methods The fraction Ⅲ of human plasma was purified by Lys Sepharose 4B affinity chromatography and Sephadex G 200 gel filtration.Results The nature plasminogen(Glu HPg)with a molecular weight of 90000 was prepared by the above mentioned method and further dialyzed,then Lys HPg was obtained.The 2 kinds of HPg were stored at different temperatures for different time,the result showed that Glu HPg was more stable than Lys HPg.Conclusion The study laid a foundation of synthesis of acylating plasminogen streptokinase complex.
2000 04 [Abstract][OnlineView][Download 50k] - Su Yan, Wang Jilin, Xue Honggang et al( Wuhan Institute of Biological Products, Wuhan 430060)
Objective To prepare biodegradable PLA/PLG microspheres used for encapsulating ribonucleoprotein(RNP)and study on the effect of it as an adjuvant.Methods Biodegradable PLA/PLG microspheres were prepared, and the quality of them were evaluated using protein loading, encapsulation rate, shape and size as indexes. The hydrolysis and release of the microspheres prepared with 3 kinds of materials were observed at 37℃ in vitro, and the immune effects of them were compared.Results At the 2nd to 4th week after immunization, the antibody titer of PLG50/50 microsphere was 2 times higher than those of PLA and PLG85/15 microspheres, and, at the 8th week, the titer of PLA microsphere reached peak value. However, all the antibody titers induced by the 3 kinds of microspheres were 5 times higher than that induced by RNP without encapsulation. The anti NP level induced by a single dose of RNP encapsulated with PLA/PLG microsphere was similar to that induced by 2 doses of RNP using Al(OH) 3 as adjuvant. However, the level was significantly higher than that induced by RNP without any adjuvant.Conclusion PLA/PLG microsphere is a good adjuvant of RNP and worth further investigation.
2000 04 [Abstract][OnlineView][Download 82k]
2000 04 [Abstract][OnlineView][Download 12k] - Luo Xuan,Wei Zili,Chen Wange et al( Changchun Institute of Biological Products,Changchun 130062 )
Objective To improve the immuogenicity of hepatitis B vaccine.Methods Mice were immunized with the hepatitis B vaccine using interferon and Al(OH) 3 complex adjuvant,and the humoral and cell immune responses of them were studied.Results Both humoral and cell immunity of the vaccine using complex adjuvant were significantly higher than that using Al(OH) 3 adjuvant alone.Conclusion Interferon can be used as an adjuvant of hapatitis B vaccine.
2000 04 [Abstract][OnlineView][Download 26k] - Wang Guangjie *,Zhai Xiaoping,Wang Gang et al( * Blood Center of Inner Mogolia,Hohhot 010010 )
Objective To establish the bank and reaction pattern of panel red cell for the clinical identification of specific antibody.Methods Twelve blood group antigens,such as ABO,Rh,MNSs and Duffy ,were screened by saline,enzyme and indirect anti human globulin methods.Results A panel red cell bank consisting of 29 kinds of blood group antigens(D,C,E,c,e,C w,M,N,S,s,P 1,Le a,Le b,K,k,Kp a,Kp b,Kp c,JS a,JS b,Fy a,Fy b,Di a,Xg a,Lu a,Lu b,Jk a,Jk b,Jr a)and 9 groups of O type Rh genetypes(R 1R 1,R 2R 2,R 1R 2,R 1r,R 2r,R 1R z,R 2R z,R 2R z,rr,rr 1)was established.On the basis of this,a reaction pattern consisting of 15 kinds of group ed cells was also established.Conclusion The probability of identifying antibodies by the 20 kinds of antigens in the bank meet related requirment(P<0.05).
2000 04 [Abstract][OnlineView][Download 45k] - Liu Changnuan,Liu Lan,,Zhang Yi et al( National Institute for the Control of Pharmaceutical and Biological Products, Beijing 100050 )
Objective To establish the quality control methods for the chemical modified interferon product.Methods Bioactivity was assayed by Wish VSV system.Molecular weight was determined by matrix asisted laser desorption ionization time of flight mass spectrometry(MALDI TOF MS).Isoelectric point was measured by high performance capillary isoelectric focusing.Peptide map was determined after trypsin treatment.Results The quality control methods were established and used for detecting 3 lots of chemical modified interferon.The specific activity,molecular weight and pI of the 3 lots were 10 6 IU/mg protein,63057 and 5.6 respectively.Conclusion The methods can be used for the quality control of chemical modified interferon product.
2000 04 [Abstract][OnlineView][Download 32k]
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2000 04 [Abstract][OnlineView][Download 19k] - Xiong Renping,Zhou Yuanguo,Liu Ping et al( The Molecular Biological Center,Research Institute of Surgery and Daping Hospital,The Third Military Medical University,Chongqing 400042 )
Objective To extract thymosin,detect the pharmaceutical indexes of it,observe the effect of it in clinical application and animal test,and explore the influence of it on the expression of Ga protein in the lung tissue of guinea pig with asthma.Methods Thymosin was prepared by acid base precipitation and centrifugation,and the pharmaceutical indexes of it were detected by quantification of polypeptide,identification of activity,bacterial culture,toxicity test and pyrogen test.The clinical effect of it was observed.Guinea pig model with asthma was established by aerosol inhalation of egg albumin.The Ga protein level in the lung tissue of guinea pig with asthma was analyzed by Western blot.Results The thymosin met the requirement of Chinese Pharmacopeia.The clinical effect of it was definite.The Gia and Gqa levels in the lung tissues of guinea pigs were significantly decreased after being treated with it (p<0 5).However,no significant change of Gsa level was observed.Conclusion The thymosin prepared by acid base precipitation and centrifugation can be applied in clinic.The curative mechanism of it might be changing the Ga protein level in lung tissue.
2000 04 [Abstract][OnlineView][Download 39k] -
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2000 04 [Abstract][OnlineView][Download 23k] 下载本期数据