Current Issue

  • Establishment of predictive model for long-term stability of rotavirus preparations

    SI Lu;GUO Min;MA Qiongliang;LIU Jiayin;FU Shuguang;CHEN Tong;LI Jialin;YU Ke;Lanzhou Institute of Biological Products Co.,Ltd.;

    Objective To screen candidate preparations of live rotavirus(RV) vaccine using stability data modeling.Methods The citric acid, sodium citrate and disodium hydrogen phosphate buffer bulk solutions were prepared, and sucrose and RV bulk solution were added after sterile filtration to prepare three RV liquid formulations: FA(5 × sucrose-1 × citric acid-1 × sodium citrate), FB(5 × sucrose-1 × citric acid-1 × disodium hydrogen phosphate) or FC(1 × sucrose), with MEM as the diluent and virus titer of 7. 5 logCCID_(50)/mL. The stability data of the three formulations were linearly extrapolated using 2-8 ℃ data extrapolation model and Arrhenius model, and compared with the actual stability curves at 2-8 ℃ for36 months to evaluate the predictive models and identify the optimal formulation.Results The two model analysis indicated that at temperatures ranging from 2 to 8 ℃, formulations FA and FB exhibited slower declines in virus titer, demonstrating significantly superior stability compared to FC. Both models confirmed the stability hierarchy as FA = FB > FC, with predictions based on medium-to-long-term data showing good agreement with 36-month real-time stability results.Conclusion This study successfully identified the more stable RV vaccine candidates FA and FB by establishing and validating two stability predictive models. These two models provide reliable tools for assessing the stability and efficiently screening RV vaccines.

    2025 12 v.38 [Abstract][OnlineView][Download 1031K]

  • Establishment and comparison of infection models of Bordetella pertussis aerosol and intratracheal nebulization in mice

    WANG Fen;GONG Beizhe;LIU Hongbo;FU Lie;YANG Miao;LEI Yu;ZENG Yun;ZHU Dewu;DUAN Kai;Wuhan Institute of Biological Products Co.,Ltd.,National Engineering Technology Research Center for Combined Vaccines,Hubei Vaccine Technology Innovation Center;

    Objective To establish two models of pertussis infection in mice through aerosol and intratracheal nebulization,and compare and evaluate the infection effects.Methods Using fermale NIH mice with five for each group, the aerosol infection model was set up with 10~8, 10~9, 10~(10)CFU/mL and initial concentration(1. 42 × 10~(11)CFU/mL) groups, and the infection was carried out by nebulization inhalation in a closed space for 30 minutes. Intratracheal nebulization infection model adopted a fixed nebulization dose of 25 ??L, including 10~3, 10~4 and 10~5 CFU/25 ??L infected bacteria groups, by inserting into the trachea for nebulization infection. On the 3 rd, 7 th, 14 th, 21 st and 28 th days after infection, three mice in each group were selected for lung specific gravity detection, and one mouse was randomly selected for pathological section analysis of lung tissue. Finally, the mice were infected by aerosol or intratracheal nebulization, and observed for 14 days to calculate the half lethal dose(LD_(50)).Results The lung specific gravity of mice significantly increased when aerosol infection occurred at a bacterial concentration of 10~(10)CFU/mL, or when nebulization infection occurred in the trachea with a bacterial count of 10~4 CFU/25 ??L or above. Lung slices showed that intratracheal nebulization infection caused more significant pulmonary inflammatory infiltration than aerosol infection, and was more correlated with infection time and infection dose.The LD_(50)of intratracheal nebulization infection was 1. 88 × 10~8 CFU/25 ??L, while the aerosol infection did not reach the lethal dose.Conclusion Both respiratory tract infections can successfully establish a pertussis mouse infection model. The intratracheal nebulization infection model was more relevant to the dose and time of infection, and can be used as a lethal model, which has the potential to detect the potency of pertussis vaccine.

    2025 12 v.38 [Abstract][OnlineView][Download 1270K]

  • Identification of key binding sites of Rift Valley fever virus envelope protein Gn to receptor

    MA Tianyi;MEI Chenchen ;SUN Binlian;GONG Rui;Jianghan University School of Medicine,Wuhan Institute of Biomedical Sciences,Hubei Province Key Laboratory of Cognitive and Emotional Disorders;

    Objective To identify the key sites of Rift Valley fever virus(RVFV) envelope protein Gn binding to the receptor,in order to provide a basis for the development of RVFV infection inhibitors.Methods The eukaryotic cell expression system was used to express RVFV envelope protein Gn, RVFV positive antibodies(antibodies 140, 144, 268, R12 and R17),low density lipoprotein receptor-related protein 1(LRP1) receptor domains(ClusterⅡ-Fc and ClusterⅣ-Fc) and six Gn mutants(T173A, Q174A, D176A, K180A, S291A and K294A). ELISA method or competitive ELISA were used to detect the binding activity of LRP1 receptor domain and RVFV Gn, the effect of RVFV positive antibody on the binding ability of LRP1 receptor domain and RVFV Gn, and the effect of Gn point mutation on its binding ability to RVFV positive antibody and LRP1 receptor domain. The inhibitory effect of LRP1 domain on RVFV infection was detected by pseudovirus neutralization assay.Results ClusterⅡ-Fc and ClusteⅣ-Fc domains of LRP1 exhibited specific binding to RVFV Gn protein. The binding ability of antibodies 268, R12 and R17 to Gn protein was dose-dependent, and no significant binding activity was detected for antibodies 140 and 144. Antibodies 268, R12 and R17 showed a certain competitive inhibitory effect on the binding ability of ClusterⅡ-Fc to Gn protein, and the competitive effect of antibody 268 was the most significant. Only antibody 268 showed competitive effect on ClusterⅣ-Fc. The binding activity of Gn mutants to RVFV positive antibodies was lower than that of Gn. Except for Q174A, the binding ability of other Gn mutants to ClusterⅡ-Fc and ClusterⅣ-Fc was all lower than that of Gn. Among them, the binding ability of T173A, K180A, S291A and K294A to ClusterⅡ-Fc was significantly lower than that of Gn(F = 15. 83, P < 0. 05), and the binding ability of K180A, S291A and K294A to ClusterⅣ-Fc was significantly lower than that of Gn(F = 8. 75, P < 0. 05).Conclusion K180, S291 and K294 are the key amino acid sites for RVFV Gn protein to mediate the interaction with the host receptor LRP1, revealing part of the molecular basis of the binding of Gn to LRP1 and laying the foundation for the development of inhibitors targeting virus adsorption.

    2025 12 v.38 [Abstract][OnlineView][Download 1177K]

  • Establishment of quality control method for anti-interleukin-5 receptor monoclonal antibody

    YANG Yalan;GUO Luyun;WU Gang;LI Meng;NI Yongbo;CUI Yongfei;YU Chuanfei;Division of Monoclonal Antibody Products, National Institutes for Food and Drug Control, NHC Key Laboratory of Research on Quality and Standardization of Biotech Products, NMPA Key Laboratory for Quality Research and Evaluation of Biological Products;

    Objective To establish a drug quality control method for anti-interleukin-5 receptor(IL-5R) monoclonal antibody, in order to provide reference for the quality control of anti-IL-5R monoclonal antibody and other monoclonal antibodies targeting cytokine receptors.Methods The monoclonal antibody was determined for the peptide mapping by reversed-phase ultra-performance liquid chromatography(RP-UPLC), for the charge variants by imaged capillary isoelectric focusing(iCIEF), for the molecular size variants by reduced/non-reduced capillary electrophoresis sodium dodecyl sulfate(CESDS) and size exclusion chromatography(SEC), for the antibody-dependent cell-mediated cytotoxicity(ADCC) activity by reporter gene assay, and for the N-glycan profile by 2-aminobenzamide(2-AB) labeling method.Results The peptide mapping of the anti-IL-5R monoclonal antibody exhibited visual similarity to the reference standard. The iCIEF main peak content was(66. 94 ± 1. 52)%. The reduced CE-SDS heavy chain plus light chain content reached(96. 39 ± 0. 11)%, while the non-reduced main peak content was(96. 98 ± 0. 13)%. The SEC monomer content was(99. 62 ± 0. 06)%. The ADCC biological activity showed a relative potency of(90. 33 ± 15. 42)%. For the N-linked glycan profile, the G1/G0 ratio was(0. 14 ± 0. 00)%, and the high-mannose content was(1. 37 ± 0. 05)%.Conclusion A comprehensive quality control methodology was established for the anti-IL-5R monoclonal antibody, encompassing identity verification, purity analysis, biological activity, and N-linked glycosylation profiling, which provides reference for the development and quality control of therapeutic antibodies against this target and other cytokine receptors.

    2025 12 v.38 [Abstract][OnlineView][Download 923K]

  • Monitoring and analysis of non-national immunization program vaccination in Liaoning Province from 2018 to 2024

    DUAN Yatong;WANG Yan;LI Meng;FANG Xing;Liaoning Provincial Center for Disease Control and Prevention;

    Objective To analyze the use of non-national immunization program(nNIP) vaccines in Liaoning Province from2018 to 2024, evaluate the current situation of nNIP vaccination in Liaoning Province, and to provide scientific basis for formulating public health policies and optimizing vaccination strategies.Methods The nNIP vaccination data from 2018 to2022 and 2023 to 2024 were extracted through “China Immunization Program Information Management System” and“Liaoning Province Immunization Program Comprehensive Information Platform-Vaccination Information System”, including nNIP vaccine replacement records. Pearson correlation coefficient matrix analysis was performed using SPSS 13.0 software,and descriptive analysis method was used to comprehensively analyze the data.Results From 2018 to 2024, a total of25 726 024 doses of nNIP vaccines were administered in Liaoning Province, with per capita dose of 866 doses per 10 000 people. The per capita doses of nNIP vaccine and the proportion in total doses of all vaccines were increasing year by year.The top three nNIP vaccine varieties used were rabies vaccine for human use(Rab), influenza vaccine(InfV) and varicella attenuated live vaccine(VarV). Among the NIP, poliomyelitis vaccine(PV) showed the highest substitution rate(30. 40%),followed by hepayitis A attenuated live vaccine(HepA-L)(29. 08%) and diphtheria,tetanus and acellular pertussis combined vaccine(DTaP)(16. 37%).Conclusion The importance of nNIP vaccines in public health has become increasingly prominent. As an important part of vaccination, the quality of surveillance needs to be further strengthened. The nNIP vaccines can make up for the lack of NIP vaccine coverage, and can also play an important role when some NIP vaccines are temporarily in short supply. On the basis of optimizing the supply and distribution of NIP vaccines, increasing the level of nNIP vaccination can effectively prevent and control the epidemic of vaccine-preventable infectious diseases.

    2025 12 v.38 [Abstract][OnlineView][Download 901K]

  • Epidemiological characteristics and etiological analysis of hand,foot and mouth disease in surrounding areas of Shijiazhuang from 2014 to 2023

    LIU Zehao;CAO Yanyan;ZHAO Xin ;GE Shengwang ;WANG Le;Children's Hospital of Hebei Province,Pediatric Research Institute,Hebei Provincial Clinical Research Center for Child Health and Disease;

    Objective To explore the epidemiological and etiological characteristics of hand, foot and mouth disease(HFMD)in the surrounding areas of Shijiazhuang from 2014 to 2023, so as to provide a basis for timely detection of changes in pathogen spectrum, further optimization of treatment plans, and taking targeted prevention and control measures in advance.Methods A retrospective analysis was conducted on the clinical data of children with HFMD who visited Hebei Children's Hospital from January 1, 2014 to December 31, 2023. PCR was used to classify and identify enterovirus 71(EV71), Coxsackievirus A group 16 strain(CA16), and enterovirus undefined(EVU). Descriptive epidemiological statistics and pathogen characteristics analysis were performed on age, gender, season, and other data.Results A total of 26 563 specimens were tested, with a positive rate of 48. 95%(13 003/26 563) and a male to female ratio of 1. 50∶1(15 950 vs.10 613), there was a statistically significant difference in gender ratios between different years(χ~2= 19. 61, P < 0. 05), and the male positive detection rate was higher than that of females(50. 69% vs. 46. 34%), with a statistically significant difference(χ~2= 48. 26,P < 0. 05). The positive rate was highest in 2016(69. 84%) and lowest in 2020(10. 64%). There was a statistically significantly difference in the positive rate of enterovirus detection between different years(χ~2= 2 648. 40, P < 0. 05).Overall, EVU type was the dominant strain of infection in this region, accounting for 75. 37% of cases, followed by EV71 type(18. 42%). After 2018, EVU type has become the absolute dominant strain with a composition rate exceeding 89%. There was a statistically significant difference in the composition of enteroviruses among different years(χ~2= 2 217. 44, P < 0. 05).HFMD occurred throughout the year, with a single peak concentrated in June to August, with a statistically significant difference in the positive detection rate among different months(χ~2= 4 050. 78, P < 0. 05). The composition of enteroviruses in different months was as follows: with CA16 having a higher composition from May to July(7. 11% to 7. 96%), EV71 having a higher composition from March to May(32. 02% to 52. 16%), and EVU having a higher composition from June to December(71. 75% to 82. 96%), with a statistically significant difference(χ~2= 541. 67, P < 0. 05). Among different age groups, the highest positive detection rate(55. 53%) was in the age group of 3-6 years and the lowest rate(33. 42%) was in the age group of older than 6 years, with a statistically significant difference(χ~2= 842. 33, P < 0. 05). The composition of different enteroviruses in different age groups was as follows: with CA16 and EV71 having the highest composition in the age group of1-3 years(8. 33%, 23. 77%), and EVU having the highest composition in the age group of older than 6 years(91. 99%), also with a statistically significant difference(χ~2= 449. 69, P < 0. 05).Conclusion EVU is a common infectious pathogen in the surrounding areas of Shijiazhuang, with a high incidence in children under 3 years old from June to August. This suggests that the epidemic trend of HFMD should be closely monitored, vaccination should be strengthened, and prevention and control measures should be strengthened for key populations.

    2025 12 v.38 [Abstract][OnlineView][Download 882K]

  • Development, verification and application of an assay for the size of host cell residual DNA fragments in pandemic influenza vaccine(H5N1)

    QIU Ran;ZHANG Qingmei;LE Yang;JI Deming ;WU Dongping;ZHANG Xiaoyu;LI Fang;WU Wenyi;YUAN Xiaoling ;ZHANG Qimeng ;LIU Bo;ZHANG Zhegang;Viral Vaccine Research and Development 2 Unit,Wuhan Institute of Biological Products Co.,Ltd.;

    Objective To develop and verify an analytical method for the size of host cell residual DNA(HCD) fragments in MDCK cells based on magnetic bead method and quantitative real-time PCR(qPCR), so as to use it to evaluate the removal effect of HCD during the production process of pandemic influenza vaccine(H5N1).Methods The HCD of samples was extracted by magnetic bead method, and specific primers and probes were designed to establish qPCR analytical method that can quantify 84, 142, 204 and 504 bp DNA fragments, respectively. The established method was verified for the linear range,limit of quantitation(LOQ),precision, accuracy, robustness and specificity, and used to analyze the proportion of HCD fragments in the process samples of pandemic influenza vaccine(H5N1), and the HCD content was simultaneously detected.Results The linear ranges of 84, 142, 204 and 504 bp standards were 0. 003-300 pg/??L, showing a good linear relationship with Ct, and the coefficients of determination(R2) of the four standard curves were all greater than 0. 99. The LOQs of 84, 142 and204 bp were 3 fg/??L, and the LOQ of 504 bp was 1 fg/??L. The recovery rates of spiked samples at high, medium and low concentrations corresponding to 84, 142, 204 and 504 bp were all within the range of 50% to 150%. The CVs of precision,robustness and specificity verification were not greater than 40%. The proportion of large HCD fragments in the harvest solution of pandemic influenza vaccine(H5N1) was relatively high(≥ 204 bp fragments accounted for 81. 0%). After enzymatic hydrolysis and purification, the proportion of ≥ 204 bp fragments in the bulk solution was reduced to 8. 9%, and no ≥ 504 bp fragments were detected. At the same time, the total HCD content was reduced from 689. 9 ng/m L to 10. 2 ng/m L.ConclusionThe q PCR analytical method for HCD fragment size was developed with good precision, accuracy, robustness and specificity,which can be used for the analysis and detection of HCD fragments of pandemic influenza vaccine(H5N1) and the quality control in the production process.

    2025 12 v.38 [Abstract][OnlineView][Download 909K]

  • Development and verification of a size exclusion-high-performance liquid chromatography method for simultaneous determination of free proteins and residual 4-dimethylaminopyridine in bulk solution of pneumococcal polysaccharide-protein conjugate

    WANG Liru;YANG Jinlong;KE Zhiping;ZHANG Yu;HU Xiaohua;Beijing Zhifei Lvzhu Biopharmaceutical Co., Ltd.;

    Objective To develop a size exclusion-high-performance liquid chromatography(SEC-HPLC) method for the simultaneous determination of free proteins and residual 4-dimethylaminopyridine(DMAP) in the bulk solution of pneumococcal polysaccharide-protein conjugate, and to verify and preliminarily apply the method.Methods By studying the characteristic ultraviolet absorption peak of DMAP and optimizing the separation method of chromatographic column, the SEC-HPLC method was established to realize the retention and separation of free proteins and DMAP on the same chromatographic column and quantify them. The method was verified for specificity(system suitability), limit of quantification(LOQ), accuracy, precision, linearity and range, and robustness, and was used to detect the free proteins and DMAP residues in the test sample.Results The chromatographic conditions were determined as follows: TSKgel G5000 PWXL analytical column was used as the column(7. 8 mm × 300 mm, 10 ??m), the column temperature was set at 35 ℃, the mobile phase was0. 85% sodium chloride solution, the pH was(6. 2 ± 0. 2), the flow rate was 0. 5 mL/min, and the detection wavelength was280 nm. The resolution of free protein peaks of diphtheria toxoid(DT) or tetanus toxoid(TT) and DMAP peak from adjacent peaks was greater than 1. 5. The LOQs of TT, DT and DMAP were 0. 50, 1. 20, and 0. 02 ??g/mL, respectively. The spiked recovery rates of TT, DT and DMAP were 98. 11%-104. 60%, 95. 43%-103. 26% and 97. 11%-98. 42%, respectively.The relative standard deviations(RSDs) of intermediate precision of TT, DT and DMAP were all less than 5%. The R~2 values of TT and DT were greater than 0. 999 0 in the range of 6-24 ??g/mL, and the R~2 value of DMAP was 1. 000 in the range of2-10 ??g/mL. The RSDs of TT, DT and DMAP content were all less than 3% at the injection temperature of 30, 35 and 40 ℃.The established method was used to determine the content of free proteins and DMAP in the test sample, and rapid and accurate results were obtained.Conclusion The established SEC-HPLC method has good system suitability, accuracy,precision, linear relationship and robustness, which can realize the dual quantification of free proteins and DMAP in the same experiment, and can be used to monitor the impurities in the binding process.

    2025 12 v.38 [Abstract][OnlineView][Download 1043K]

  • Preparation of monoclonal antibody against ovalbumin and establishment and verification of double antibody sandwich ELISA for quantitative detection

    YU Boshan;HU Weiyan;SUN Ziqi;WU Tong;DU Mo;CHANG Junliang;Changchun Institute of Biological Products Co.,Ltd.;

    Objective To prepare mouse monoclonal antibodies(mAbs) against ovalbumin(OVA), and to establish and verify a double antibody sandwich ELISA quantitative detection method for the detection of OVA residues in influenza virus split vaccine.Methods Using OVA standard as immunogen, three positive hybridoma cell lines were screened by mouse hybridoma fusion technique. The OVA mAb ascites was prepared by in vivo induction method, and then purified by ammonium sulfate precipitation and Protein A affinity chromatography. The specificity of mAb was detected by Western blot, and the relative affinity was determined by indirect ELISA. Using the overlapping coefficient as the index, the optimal paired mAbs of three hybridoma cell lines were screened, and the OVA double antibody sandwich ELISA detection method was established. The concentration of coating antibody(8. 0, 4. 0, 2. 0, 1. 0, 0. 5 ??g/mL) and the dilution of detection antibody(1∶100, 1∶500, 1∶1 000, 1∶2 000) were optimized by checkerboard titration, and then the limit of quantitation(LOQ),linear range, specificity, accuracy, precision, applicability and stability of the method were verified. The OVA content of seven batches of influenza virus split vaccine bulk solutions was determined by the established method and imported OVA detection kit, and the coincidence rate was calculated.Results A total of three hybridoma cell lines that could be stably passaged and produce OVA-specific mAb were selected, named 3D6, 2B11 and 3H8, respectively, and the ascites titers were all higher than 10~7. After purification, the purity was over 95%, with good specificity, and the relative affinity ranking was2B11 > 3D6 > 3H8. After pairing screening and optimization, it was determined that 2B11 was used as the coating antibody with the optimal concentration of 4 ??g/mL, and biotin-labeled 3D6 was used as the detection antibody with the optimal dilution of 1∶100. The method had a limit of quantitation(LOQ) of 3 ng/mL. In the concentration range of 3. 13-50. 00 ng/mL,OVA standard exhibited a good linear relationship with A_(450/630), and the linear regression equation was y = 0. 004 9 x + 0. 088 7,R~2= 0. 987 9. This method had no cross-reaction with fetal bovine serum and human serum albumin. The spiked recovery rates of three influenza virus split vaccine bulk solutions were all within the range of 90% to 110%. The CVs of precision verification were less than 10%. The dilution of three batches of influenza virus split vaccine bulk solutions showed a good dose-dependent relationship with A_(450/630). Compared with the detection reagents stored at 4 ℃ for 6 days, the detection results of the detection reagents stored at 37 ℃ for 3 and 6 days decreased by less than 20%, and each R~2 was more than 0. 97.The coincidence rates between the detection results of the established method and imported OVA detection kit were between93. 96% and 100. 89%.Conclusion The OVA double antibody sandwich ELISA quantitative detection method was established by using the prepared OVA mAb, with good specificity, accuracy, precision, applicability and stability, which provides a reliable detection method for the determination of OVA residues in the verification requirements of chicken embryo-derived vaccine finished products.

    2025 12 v.38 [Abstract][OnlineView][Download 991K]

  • Preparation of high-purity tetanus antitoxin via immunoaffinity chromatography and its application

    YAO Xiaodong;GAO Xingyue ;LIU Jiao ;XU Kai ;HU Jing ;XIAO Cong ;WANG Denghai ;LI Changqing ;JI Chong;WANG Peng ;JING Yue;Jiangxi Biological Products Institute Co.,Ltd.;

    Objective To selectively purify antibodies with high specificity for the C subunit of tetanus toxin(TT) from traditional tetanus antitoxins(TATs).Methods In this study, NHS-activated Sepharose 4 FF resin was used in combination with the TT TeNT-Hc-C869A subunit as a ligand to develop an immunoaffinity chromatography method. The stability and durability of the resin were tested by storing it under different conditions(at 4, 25 and 37 ℃ for 3, 10 and 30 d) and monitoring its reusability. The changes in the potency of the purified fractions by immunoaffinity chromatography were evaluated both in vitro and in vivo.Results Storage at 4 and 25 ℃ for up to 30 d did not significantly affect column performance. F(ab') 2 was purified by immunoaffinity column for five times continuously, and the loading capacity, elution efficiency and purity of eluted antibody were consistent each time, with the protein purity greater than 95%. After five times of continuous alkaline washing and the following purification, the purity of the protein was above 95%. The specific activities of purified equine plasma, enzymatically digested equine tetanus immunoglobulin(TIG) F(ab')2 and human TIG increased by5, 5 and 30 times respectively. And in the mouse neutralization assay, the specific activity of F(ab')2 increased significantly after purification, reaching 243 902 IU/g.Conclusion The developed immunoaffinity chromatography column shows broad applicability, and the chromatography matrix used is simple, cost-effective, and easy to replicate, making it suitable for largescale industrial applications, greatly enhancing safety, and reducing the risk of common clinical allergic reactions.

    2025 12 v.38 [Abstract][OnlineView][Download 927K]

  • Optimization of expression conditions of modified tissue plasminogen activator

    YANG Ziwei;TIAN Guanfang;CHEN Yuewen;WEN Ruixin;MIAO Yu;ZHAO Tianyu;LIANG Shimin;YAN Fengying;WU Weidang;State Key Laboratory of Druggability Evaluation and Systematic Translational Medicine,Tianjin Institute of Pharmaceutical Research;

    Objective To optimize the expression conditions of modified tissue plasminogen activator(t-PA) in order to enhance the yield and quality of modified t-PA(mt-PA).Methods The fed-batch cultivation method was used to detect the effects of low temperature culture(37 ℃ for 5 days and cooled to 30 ℃) and different concentrations of sodium butyrate(0,0. 5, 1, 2, 5 mmol/L) on the growth status of stable Chinese hamster ovarian cancer mt-PA cell line(CHOZN-mt-PA), the concentration of key metabolites(glucose, lactic acid, and ammonia) of the cells, the expression level of mt-PA, and the ratio of single/double chains. The thrombolytic activity of mt-PA in vitro was measured by bubble method.Results The cell proliferation was inhibited, the glucose consumption as well as the accumulation of lactic acid and ammonia were reduced, the expression level of mt-PA greatly increased, and the ratio of single/double chains was stable with the addition of 2 mmol/L sodium butyrate and 30 ℃ low-temperature cultivation. Compared with the corresponding control group, there was no statistically significant difference in thrombolytic activity of mt-PA prepared at 30 ℃ with the addition of 2 mmol/L sodium butyrate in vitro(t = 0. 546 32 and 0. 441 70, P = 0. 613 9 and 0. 681 5, respectively).Conclusion Cultivation at 30 ℃ and the addition of 2 mmol/L sodium butyrate can increase the expression level of mt-PA without reducing its thrombolytic activity in vitro,which provides a reliable strategy for the development of high-yield culture process of mt-PA.

    2025 12 v.38 [Abstract][OnlineView][Download 1025K]

  • Establishment and verification of a method for determination of aluminum content in vaccines by high performance liquid chromatography-diode array detection with 8-hydroxyquinoline pre-column derivatization

    LIU Yijie;FU Honghong;WANG Xinjin;WU Jingping;WANG Xin;CHEN Liang;ZHENG Wenjing ;YE Yanyan;Fujian Institute for Food and Drug Quality Control;

    Objective To establish an 8-hydroxyquinoline pre-column derivatization-high performance liquid chromatographydiode array detection(HPLC-DAD) method for the determination of aluminum content in vaccines, and to verify the method for vaccine quality control.Methods The determination was carried out by pre-column derivatization with 8-hydroxyquinoline, and the sample pretreatment conditions of hydrochloric acid solution concentration(0. 1, 0. 5, 1, 2, 5 mol/L),hydrochloric acid digestion time(20, 30, 40, 50, 60 min), concentratio%n of derivatization reagent 8-hydroxyquinoline(0. 5,1, 2, 3, 4 mg/mL), and ammonia concentration(1. 25%, 6. 25%, 12. 50%, 18. 75%, 25%) were optimized. ZORBAX SB-C_8(4. 6 mm × 150 mm, 5 ??m) column was used with methanol-water as mobile phase in gradient elution at a flow rate of0. 8 mL/min. The detection wavelength was 254 nm, the column temperature was 30 ℃, and the injection volume was 10 ??L.The linear range, accuracy, precision, specificity and stability of the method were validated, the limit of detection(LOD) and limit of quantification(LOQ) were determined, and the method was used to detect the aluminum content in 16 batches of bivalent human papillomavirus(HPV) and three batches of recombinant hepatitis E virus(HEV) vaccines.Results The optimized pretreatment conditions were as follows: hydrochloric acid concentration 1 mol/L, digestion time 20 min,derivatization reagent 8-hydroxyquinoline concentration 2 mg/mL, and ammonia concentration 12. 5%. Aluminum showed a good linearity in the concentration range of 2 to 100 ??g/mL, with a correlation coefficient(r) of 0. 999 98. The average recovery rates of spiked samples were between 89. 6% and 95. 7%. The relative standard deviation(RSD) of the aluminum content test results of six samples was 1. 4%. The blank reagent showed no interference with the detection. The sample solution exhibited good stability at room temperature for 12 h. The LOD and LOQ of the method were 0. 267 ??g/mL and0. 820 ??g/mL respectively. The established method was used to detect the aluminum content in 16 batches of HPV vaccines and three batches of recombinant HEV vaccines, and the results were not significantly different from those detected by titration method in General Principles 3106 of the Chinese Pharmacopoeia(Volume Ⅲ, 2020 edition).Conclusion The established HPLC-DAD method is accurate and reliable, with good precision, specificity and stability. The mobile phase is only methanol-water without adding buffer salts and derivatizing agents, etc., and it is easy to operate and friendly to instruments and chromatographic columns. The method requires a small amount of samples, which can realize the detection of a single dose of vaccine sample and provide reference for the quality control of vaccines.

    2025 12 v.38 [Abstract][OnlineView][Download 922K]

  • Optimization of clarifying filtration and ultrafiltration concentration process for harvest solution of Coxackievirus A16

    LU Xiaowei;ZHENG Zhen;CAO Mingxiang;XU Hua;LI Sihao;ZHANG Haoran;YANG Erxia;Aimei Action Biopharmaceutical Co.,Ltd.;

    Objective To investigate a technical approach suitable for the clarification and concentration of Coxackievirus A16(CA16) harvest solution, aiming to effectively remove impurity particles and proteins while achieving high antigen concentration.Methods The virus harvest solution underwent two-stage filtration pretreatment. Antigen recovery rate,membrane flux, and turbidity were utilized as evaluation metrics at each stage to examine the effects of various membrane materials, pore sizes, and structures on clarifying efficacy. Subsequently, clarified harvest liquid was concentrated using a100 KD polyether sulfone membrane, and the changes in antigen content and protein content were analyzed to determine the optimal concentration multiples, washing cycles, and sampling intervals. The sample size was scaled up for pilot testing where the harvested liquid was clarified, filtered and subjected to ultrafiltration based on the optimized parameters. The linear scalability of this process was assessed using antigen recovery rates and protein removal rates as indicators.Results In the first stage, 0. 45 ??m glass fiber membranes were employed while in the second stage 0. 45 ??m polyether sulfone membranes were used. The optimal conditions for ultrafiltration concentration included a concentration factor ranging from 30 to50 times with four washing filtrations followed by four sampling collections. At pilot scale testing, post-clarification resulted in a turbidity reduction of CA16 virus by 96. 00%, with an antigen recovery rate of 89. 49%. Following ultrafiltration concentration yielded an antigen recovery rate of 87. 23% alongside a protein removal rate reaching 91. 17%.Conclusion The optimized ultrafiltration concentration technology not only reduces protein content but also achieves high levels of antigen recovery while demonstrating excellent potential for linear amplification.

    2025 12 v.38 [Abstract][OnlineView][Download 860K]

  • Molecular mechanism of cerebellin family regulating synapse formation and research progress on it in prevention and treatment of neurological diseases

    DU Jinfeng;YANG Jian ;ZHAO Panfeng;Shanghai GeneoDx Biotech Co., Ltd.;

    The development of the nervous system and functional recovery following injury critically depend on synaptic involvement, with synaptic plasticity regulation serving as the foundational mechanism underpinning neural development,functional maintenance, and repair. The cerebellin(Cbln) protein family, widely distributed synaptic organizers in the brain,mediates specific interactions between secreted proteins and receptors to drive synapse assembly, maintenance, and neural circuit formation, thereby achieving synaptic plasticity modulation. Notably, dysfunction of this family is closely associated with multiple neurological disorders. This paper reviews the mechanistic roles of Cbln family members in synaptic molecular regulation, signal transduction, and neural network construction, providing comprehensive and detailed clues for the study of cerebellin family and valuable research perspectives for the prevention and treatment of related neurological diseases.

    2025 12 v.38 [Abstract][OnlineView][Download 993K]

  • Progress in research on group B Neisseria meningitidis vaccine

    GU Tiejun;GU Hanxue ;XI Hualong ;SUN Bo;School of Life Science,Jilin University;

    Neisseria meningitidis(Nm) is the primary pathogen causing epidemic cerebrospinal meningitis, which can be classified into 13 serogroups based on its capsular polysaccharides. With the successful development and widespread use of meningococcal vaccines targeting serogroups A, C, Y, and W135, group B Neisseria meningitidis(MenB) has gradually become one of the main pathogens responsible for invasive meningococcal disease(IMD). This paper reviews the commercially available MenB vaccines overseas and the recent discovery of potential MenB antigens, aiming to provide reference for MenB vaccine research and development.

    2025 12 v.38 [Abstract][OnlineView][Download 899K]

  • Study on pathogenesis and new therapy of enterovirus-associated autoimmunity diseases

    JIANG Yilin;LI Jiakai;AN Qixv;ZHENG Baisong;Central Laboratory, Lequn Campus, First Hospital of Jilin University;

    Autoimmunity diseases(AIDs) involve the entire body's tissue systems and pose a serious threat to the normal immune system and health of the human body. Researches have shown that AID is caused by the complex interaction between genetic susceptibility and environmental factors. Enterovirus(EV) infection often accompanies certain AID and is an important type of virus infection related to AID. Coxsackievirus(CV), a member of the EV family, is an important pathogen among the pathogenic factors of type 1 diabetes(T1D). EV infection causes damage to the activity of pancreatic β cells and accelerates the disease progression of T1D. In terms of treatment, recent reports have shown that antiviral therapy for T1D is effective in preserving the function of β pancreatic cells. In celiac disease and Crohn's disease(CD), EV infection can induce inflammatory pathogenic changes in the intestinal microbiota, increase intestinal barrier permeability, and may trigger the occurrence of such AIDs. However, the relationship and mechanism between EV infection and many AIDs remain unclear.This paper summarizes the research conclusions on the correlation between EV infection and AIDs to facilitate a deeper exploration of their pathogenic patterns.

    2025 12 v.38 [Abstract][OnlineView][Download 911K]

  • Research progress on quality control of glycoprotein biosimilars

    ZHOU Zhongnan;CAI Na;FENG Rui;MA Taotao;School of Pharmacy, Anhui Medical University;

    Biological products are a class of complex macromolecular drugs produced by biological systems(such as microorganisms and cell lines) and prepared by biotechnology methods, such as monoclonal antibodies, fusion proteins, vaccines,interferons and growth hormone. Biosimilar drugs are highly similar in structure and function to the original drugs, which can not only significantly reduce medical costs, but also significantly improve patients' accessibility to high-quality drugs, thus having broad development prospects. Common biosimilar modification methods include glycosylation, lipidation, nucleic acid modification, etc. Glycosylation is a process of modification of translated proteins that can affect the physical and chemical characteristics of therapeutic proteins such as safety, solubility, and stability, which is one of the critical quality attributes(CQAs) of proteins. This paper reviews the research progress on quality control of glycoprotein biosimilars, so as to provide reference for the related research and development in the future.

    2025 12 v.38 [Abstract][OnlineView][Download 897K]

  • Construction of a quality management system for entire life cycle of vaccine pharmaceutical development and analysis of key inspection points

    MA Yansong;KANG Ying;DONG Fang;ZHAO Xin;Center for Food and Drug Inspection of NMPA;

    Pharmaceutical research and development of vaccines is the core link to ensure their successful transition from laboratory research to commercial stable production. In view of the fact that my country's current vaccine development still mostly draws on traditional processes, the construction of the quality management system(QMS) for the entire life cycle of pharmaceutical development is not yet perfect, and there are challenges such as data reliability management, it is very important to establish a scientific and effective quality management strategy. Based on a systematic analysis of international and domestic regulatory guidelines, this paper proposes to establish a phased and differentiated quality management system, and emphasizes the concept of Quality by Design(QbD) as the core guide. The application of this system in the entire life cycle of vaccine development is systematically expounded, covering the key control points such as seed lot management for bacterial and viral strains, manufacturing process development and optimization, and quality research, as well as quality management strategies at each stage, in order to provide systematic reference for improving the compliance and reliability of vaccine development and meeting the regulatory expectations, so as to ultimately ensure the quality and safety of marketed vaccines.

    2025 12 v.38 [Abstract][OnlineView][Download 900K]


@2012 Editorial Department of "Chinese Journal of Biologicals"