2025年04期目次

  • Optimization of culture process and characterization of critical quality attributes of Coxsackievirus A16

    WAN Xin;XIAO Ao;WANG Wenhui;CAI Qian;MENG Shengli;GUO Jing;WANG Zejun;SHEN Shuo;Viral Vaccine Research Laboratory, Wuhan Institute of Biological Products Co., Ltd., National Engineering Technology Research Center of Combined Vaccines, Vaccine Technology Innovation Center of Hubei Province;

    Objective To optimize the culture process of Coxsackievirus A16(CV-A16) and compare the different quality attributes of CV-A16 harvest solution in order to determine the critical quality attributes(CQAs), and lay a foundation for the development of the preparation process and quality control method of CV-A16 vaccine. Methods Using Vero cells as matrix,virus titer and mature complete viral particle antigen content(1H5 antigen) as evaluation indicators, the cell growth phase(mid-log phase, late-log phase, plateau phase), MOI(0. 01, 0. 001, 0. 000 1), culture medium type(serum-free M199, DMEM,50% M199 + 50% DMEM medium), serum concentration of culture medium(serum-free, 1%, and 2% newborn bovine serum) and culture temperature(33, 35, and 37 ℃) for CV-A16 inoculation were optimized, and the optimized process was further verified in T25 cell bottle and 10-layer cell factory. Correlation analysis was performed between virus titer, antigen contents of mature complete viral particles(1H5 and 20E8 antigens), and neutralizing epitope antigen content(3H4 antigen)with immunogenicity to identify critical quality attributes(CQAs). Results The optimum culture conditions of CV-A16 were determined by single factor experiment and enlarged scale culture. Vero cells were inoculated at a MOI of 0. 001 at the end of logarithmic phase and cultured at 33 ℃ in serum-free M199 medium. The antigen contents(1H5 and 20E8 antigens) of mature complete viral particles obtained from CV-A16 harvested solution under four culture conditions(serum-free DMEM culture medium at 33 ℃ for 6 d, serum-free M199 culture medium at 33 ℃ for 6 d, serum-free DMEM culture medium at37 ℃ for 4 d, serum-free M199 culture medium at 37 ℃ for 4 d) were quite different, and the trend of change was different from that of neutralizing antibody titer. The virus titer and neutralizing epitope antigen content(3H4 antigen) of CV-A16harvested solution obtained under the four process conditions changed little, and the trend of change was the same as that of neutralizing antibody titer. Conclusion Based on the quality attribute of immunogenicity(neutralizing antibody titer), the virus titer and neutralizing epitope antigen content(3H4 antigen) can be classified as the CQA of CV-A16 virus harvest solution,and the antigen contents of mature complete viral particles(1H5 and 20E8 antigens) can be used for the analysis of virus particles in the quality control process of CV-A16 production.

    2025 04 v.38 [Abstract][OnlineView][Download 1031K]

  • Immune effect of inactivated foot-and-mouth disease vaccine combined with goat pox vaccine and live Brucella vaccine

    LI Xiaozhuo;JIN Yinghong ;ZHANG Ling;LI Yue ;WANG Ping ;LIU Wenkai ;HAN Xiangshu ;LIANG Xianxian;ZHENG Qiming;FENG Jihong ;WANG Yan ;XIA Jun;College of Veterinary Medicine, Xinjiang Agricultural University;

    Objective To analyze the immune effect of combined use of inactivated foot-and-mouth disease vaccine(IFMDV),goat pox vaccine(GPV) and live Brucella vaccine(LBV), and to provide a new and efficient immunization model for the prevention and treatment of bovine diseases in China.Methods Twenty healthy calves were immunized with IFMDV and GPV to observe the adverse reactions after immunization. The antibody levels against foot-and-mouth disease and bovine nodular skin disease in serum were determined by liquid phase blocking ELISA and competitive ELISA respectively. Twenty healthy calves were immunized with IFMDV and LBV to observe the adverse reactions after immunization, and the serum antibody levels against foot-and-mouth disease and Brucella were determined by liquid phase blocking ELISA and tube agglutination test respectively. Finally, IFMDV, LBV and GPV were combined to immunize twenty healthy calves, and the adverse reactions of the immunized calves were observed. The serum antibody levels against foot-and-mouth disease,Brucella and bovine nodular skin disease were determined by liquid phase blocking ELISA, tube agglutination test and competitive ELISA respectively.Results No serious adverse reactions were observed in IFMDV combined with GPV or LBV immunized calves. The level of GPV antibody in serum of calves immunized with IFMDV and GPV at the same time was significantly higher than that of calves immunized with GPV alone. The level of LBV antibody in serum of calves immunized with IFMDV and LBV at the same time was higher than that of calves immunized with LBV alone. In the combined immunization group with IFMDV, GPV and LBV, the antibody levels of GPV and LBV in serum of calves were both higher than those of calves with single immunization.Conclusion IFMDV combined with GPV and LBV has good safety, and can improve the immune effect of GPV and LBV.

    2025 04 v.38 [Abstract][OnlineView][Download 940K]

  • Effects of ubiquitin-like modifier activating enzyme 1 on proliferation and apoptosis of acute myeloid leukemia cells and its molecular mechanism

    ZHANG Aili;ZHAI Li;ZHANG Yongrui;GAO Yanzhang;PAN Yuqing ;GUO Chengbin;LU Yanzhou;YANG Wei;ZHANG Xi;Department of Clinical Laboratory, Peking University Cancer Hospital Yunnan, Caner Hospital of Yunnan Province, Third Affiliated Hospital of Kunming Medical University;

    Objective To investigate the effects of ubiquitin-like modifier activating enzyme 1(UBA1) on the proliferation and apoptosis of acute myeloid leukemia(AML) cells(HL60 and THP-1 cells) and its mechanism, so as to evaluate the possibility of using UBA1 as a molecular marker and target for diagnosis and treatment of AML. Methods The expression of UBA1 in HL60 and THP-1 cells was inhibited via shRNA interference, and stable transfection cell lines were screened. The inhibition effect was detected by qPCR and Western blot. The effect of inhibiting UBA1 on proliferation of AML cells was measured by CCK-8 assay. Flow cytometry was used to detect the effect of inhibiting UBA1 on apoptosis of AML cells. The effects on the expression of apoptosis proteins(Bax, Bc12), cell cycle regulation related proteins(CDK4, CDK6 and CyclinD1) and MAPK signaling pathway related proteins(P-ERK, P-JNK, P-P38MAPK, T-ERK, T-P38) in AML cells were determined by Western blot. Results Following UBA1 knockdown, both HL60 and THP-1 cells exhibited a significant reduction in UBA1 mRNA transcription and protein expression levels(t = 2. 065-43. 591, each P < 0. 05). Cell proliferation capacity was significantly suppressed(t = 12. 274-17. 252, each P < 0. 05), while the apoptosis rate increased significantly(t = 12. 690-18. 855, P <0. 05). The pro-apoptotic protein Bax was significantly upregulated(t = 17. 094-20. 781, P < 0. 01), whereas the anti-apoptotic protein Bcl-2 was downregulated(t = 42. 494-53. 050, P < 0. 01). The expression levels of cell cycle regulatory proteins CDK4, CDK6, and Cyclin D1 all significantly decreased(t = 12. 193-51. 666, each P < 0. 05). The expression levels of P-ERK and P-P38MAPK in MAPK signaling pathway increased significantly(t = 3. 759-10. 822, each P < 0. 05),but the expression levels of P-JNK, T-ERK and T-P38 had no statistically significant difference(t = 0. 133-1. 794, each P >0. 05). Conclusion Interference with the expression of UBA1 can inhibit the proliferation of HL60 and THP-1 cells and promote their apoptosis, of which the mechanism may be related to the increased expression of P-ERK and P-P38MAPK proteins in MAPK signaling pathway.

    2025 04 v.38 [Abstract][OnlineView][Download 1381K]

  • Comparative analysis of inflammatory responses in lung tissue of K18-hACE2 transgenic mice infected with SARS-CoV-2 prototype and variant strains

    SUN Kaili;XU Xiuli;LI Chunyang;LU Jia;WANG Zejun;Wuhan Institute of Biological Products Co., Ltd.;

    Objective To compare the immune responses in lung tissue of K18-hACE2 transgenic mice infected with SARS-CoV-2 prototype strain Prototype and its variants Delta and Omicron BF.7, so as to provide experimental evidence for the research and development of related drugs. Methods K18-hACE2 transgenic mice were infected with Prototype, Delta or Omicron BF.7 by nasal drip. On the 7th day after infection, the lung tissues of mice were taken aseptically for subsequent analyses. Viral RNA copy numbers and cytokine gene expression levels in the lung tissues were quantified by qPCR. Histopathological examination of lung tissue damage was performed using HE staining. Results The copy number of viral nucleic acid in lung tissue of mice in Delta group was significantly higher than that in Prototype and Omicron BF.7 groups(t = 5. 419-6. 908, each P < 0. 01), the expression levels of interferon γ(IFN γ)and interleukin-13(IL-13)in lung tissue were significantly higher than those in Prototype and Omicron BF.7 groups(t = 3. 990-7. 282, each P < 0. 05), and the expression levels of tumor necrosis factor-α(TNF-α),and IL-5 were significantly higher than those in Prototype group(t = 7. 108 and 2. 908,each P < 0. 05). In addition, obvious pathological changes related to virus infection were observed in lung tissue sections of mice in three groups. Conclusion Delta variant strain induced the strongest inflammatory response in lung tissue of K18-hACE2 transgenic mice with the highest pathogenicity, followed by prototype strain and Omicron BF.7 variant strain.

    2025 04 v.38 [Abstract][OnlineView][Download 1215K]

  • Extraction, identification, and immunogenicity analysis of Trueperella pyogenes heparin high affinity proteins

    XU Dengfeng;ZHANG Suhui;YU Yuandi;YANG Huan ;ZHAO Ziliang ;SHEN Kefei;Chongqing Academy of Animal Sciences;

    Objective To extract and identify Trueperella pyogenes(T.pyogenes) heparin high affinity proteins(THHAPs),and analyze their immunogenicity, so as to provide a reference for exploring the interaction between T.pyogenes and heparin.Methods The surface proteins of T.pyogenes were extracted using a bacterial membrane protein extraction kit. THHAPs were extracted from the surface proteins by adsorbing with heparin-agarose, washing with 1 mol/L NaCl solution, and eluting with 8 mol/L urea solution, which were then analyzed using SDS-PAGE and liquid chromatography-tandem mass spectrometry(LC-MS/MS). Bioinformatics algorithms were used to analyze the subcellular localization, signal peptides, transmembrane domains and immunogenicity of THHAPs. Fifteen female Kunming mice were immunized with THHAPs. After two weeks of the second immunization, five mice were collected for blood samples from the orbital vein and the serum was isolated, and the other 10 mice were intraperitoneally injected with 9 × 108CFU of T.pyogenes. The immunoprotective effect and antiserum hemolytic inhibitory activity were detected.Results THHAPs were observed to form a major band accounting for 98% of the protein content. A total of 1 767 pyolysin(PLO) peptides were detected, accounting for 98. 06% of the detected peptides(1767/1 802). THHAPs were composed of 1 cell wall protein, 11 secreted proteins, 1 lipoprotein, 7 membrane proteins, and 1cytoplasmic protein, 8 proteins among which might be antigens with immunoprotective properties. The mice(9/10) immunized with THHAPs were able to resist T. pyogenes infection, and their antisera exhibited hemolytic inhibitory activity.Conclusion THHAPs have been successfully extracted from the surface proteins of T. pyogenes, with PLO as their major component, which can induce a protective immune response in mice.

    2025 04 v.38 [Abstract][OnlineView][Download 938K]

  • Establishment and application of plasmid reference for detection of residual DNA of early region 1A protein and simian virus 40 large T antigen

    BI Hua;WEN Fenfen ;WANG Jiawei ;YANG Ling ;WANG Yuzhe ;QIN Xi;LIANG Chenggang;National Institutes for Food and Drug Control;

    Objective To construct a plasmid reference for the detection of residual DNA from early region 1A(E1A) protein and simian virus 40 large T antigen(SV40LTA), and to explore a digital PCR(dPCR)-based analytical technique for copy number calibration of plasmid reference to quantify plasmid reference accurately, thereby providing a new idea for host cell DNA residual detection in biological products.Methods The plasmid pUC19-E1A-SV40LTA was constructed, and the whole gene was synthesized and then amplified to obtain sufficient reference plasmid. The digital PCR system was employed to determine the copy numbers using double fluorescence channels of E1A and SV40LTA primer-probe sets, respectively.The reference material calibrated by digital PCR was subsequently applied to qPCR to establish a method for detecting the specific E1A and SV40LTA sequences. The developed qPCR assay was systematically validated for plasmid reference linearity, accuracy, limit of quantification(LOQ), precision, specificity, applicability, and stability. Furthermore, the established qPCR system was utilized for the detection of recombinant adenovirus(rAdV) sample and recombinant adeno-associated virus(rAAV) sample.Results The copy numbers of E1A and SV40LTA targets in the plasmid reference were found to be essentially consistent through digital PCR using dual fluorescence channels. The detection values detected by E1A and SV40LTA primer-probe sets were 3. 55 × 109and 3. 48 × 109copies/μL, respectively. The coefficient of variation(CV) of these two values was 1. 42%, and the mean value of 3. 51 × 109copies/μL was taken as the calibration value of digital PCR.The qPCR system established by using digital PCR to calibrate copy number exhibited good linearity, accuracy, LOQ, precision, specificity, applicability and stability, and the recovery rates for rAdV and rAAV samples were between 70% and130%.Conclusion A reference plasmid for detecting E1A and SV40LTA residues in biological products was established,and a more sensitive and accurate digital PCR method was introduced for quantification, which can be used to detect the residual DNA of E1A and SV40LTA in gene therapy products produced with HEK293 and HEK293T as host cells.

    2025 04 v.38 [Abstract][OnlineView][Download 984K]

  • Expression design and optimization of full-length monoclonal antibodies in Pichia pastoris

    YE Kaixiong;GONG Xiulong;XU Mingqiang;QIAN Zhilan;LIU Qi;CAI Menghao;State Key Laboratory of Bioreactor Engineering, School of Bioengineering, East China University of Science & Technology;

    Objective To express monoclonal antibodies(mAbs) in Pichia pastoris chassis host and explore various expression optimization strategies to improve the yield of antibodies.Methods Pichia pastoriswas utilized as the chassis cell for the recombinant expression of various mAbs using both methanol-induced system PAOX1 and ethanol-induced system E1.Molecular chaperone overexpression and media component optimization were employed to further enhance the production of mAbs. The productivity of the recombinant strains was then evaluated at a 3 L reactor scale.Results Ethanol-induced expression of Eptinezumab resulted in a yield of 12. 4 mg/L. Overexpression of molecular chaperones Ppi1 and Biph increased Eptinezumab production by approximately 22. 5% and 23. 0%, respectively. Additionally, the supplementation of1. 0 g/L ammonium sulfate, the adjustment of 0. 05 mol/L potassium phosphate buffer, and the addition of 80 mmol/L threonine individually enhanced Eptinezumab yields by approximately 57. 1%, 25. 8%, and 58. 1%, respectively. At the 3 L bioreactor scale, the Biph-overexpressing strain exhibited a remarkable increase in complete antibody production, achieving a maximum yield of 74. 1 mg/L, which was approximately 5. 9 times higher than that obtained at the flask level.Conclusion Eptinezumab can be successfully expressed in the Pichia pastoris chassis host, and the application of various expression optimization strategies has effectively enhanced the production of the antibody. This study provides novel insights and directions for the expression of monoclonal antibodies in Pichia pastoris.

    2025 04 v.38 [Abstract][OnlineView][Download 1010K]

  • Genetic characteristics of viruses related to viral diarrhea in Jilin Province

    LI Xiang;CUI Fangjian;XU Shuang;MENG Tingyu ;CHENG Tao;WANG Shuang;WANG Ao;HUANG Biao;YANG Yao;Jilin Provincial Center for Disease Control and Prevention, Jilin Provincial Academy of Preventive Medicine;

    Objective To investigate the genetic characteristics of viruses associated with viral diarrhea in Jilin Province, in order to provide a scientific basis for making more effective prevention and control strategies of viral diarrhea.Methods A total of 211 fecal specimens from hospitalized children under 5 years of age with diarrhea and 7 sewage samples were collected in Jilin Province from January to July 2023. Initial screening was performed using qPCR, and positive specimens were further analyzed by RT-PCR to amplify target genes. Sequencing was conducted, and phylogenetic trees were constructed to analyze genetic relationships.Results In fecal specimens, the positive rates for rotavirus(RV), Norovirus(NoV),Sapovirus(SaV), astrovirus(AstV), and adenovirus(AdV) were 10. 9%, 48. 3%, 4. 3%, 1. 4%, and 5. 7%, respectively. In sewage samples, 5 AstV and 4 NoV were detected, along with 3 samples each of RV, SaV, and AdV. Sequencing and genotyping results revealed that RV G8P [8] was the predominant circulating strain. Among NoV strains, GⅡ.4 [P16] was the most prevalent, followed by GⅡ.4 [P31]. In fecal samples, GⅠ group strains included GⅠ.3 [P13] and GⅠ.1 [P1]. One SaV strain of GⅡ.5 was identified. For AstV, three strains of HAstV-1a and one strain of HAstV-5 were detected. Among AdV strains, three were identified as HAdV-41, three as HAdV-2, one as HAdV-1, and one as HAdV-5.Conclusion The pathogenic genes of viral diarrhea in Jilin Province are diverse, the dominant strains of RV have changed, and domestic sewage contains a variety of diarrhea viruses.

    2025 04 v.38 [Abstract][OnlineView][Download 1127K]

  • Establishment and validation of droplet digital PCR assay for detection of template DNA residues in SARS-CoV-2 mRNA vaccine

    ZHAO Danhua;LIU Xinyu;HUANG Yanqiu;SUO Yue;LI Yuhua;WU Xiaohong;Division of Arboviral Vaccines, National Institutes for Food and Drug Control;

    Objective To establish and validate a droplet digital PCR(ddPCR) method for the detection of template DNA residues in SARS-CoV-2 mRNA vaccine, thereby providing technical support for the quality control of SARS-CoV-2 mRNA vaccine.Methods With the replicon of the common DNA sequence of different plasmids as the target gene, specific primers and probe were designed to establish a ddPCR method for detecting the template DNA residues in SARS-CoV-2 mRNA vaccine. The established method was validated for linear range, repeatability, accuracy, intermediate precision, robustness,limit of detection(LOD), limit of quantification(LOQ), and specificity. The validated ddPCR method was subsequently applied to detect template DNA residues in SARS-CoV-2 mRNA vaccines targeting various variants from different manufacturers.Results The optimal annealing temperature for the established ddPCR was determined to be 58 ℃. The method demonstrated good linearity within a copy number range of 19 to 4 729 copies/μL, with a correlation coefficient(R2) of 0. 999 9.The repeatability coefficient of variation(CV) was less than 10%, and the accuracy recovery rate ranged from 99% to 121%,with a CV of less than 10%. Both CVs of intermediate precision and robustness exhibited less than 10%. The LOD and LOQ of the method were 9 copies/μL and 26 copies/μL, respectively, with good specificity. When applied to detect template DNA residues in SARS-CoV-2 mRNA vaccines from four different manufacturers targeting various variants, the CVs were all not more than 15%, and the recovery rates ranged from 79% to 138%.Conclusion The established ddPCR method for detecting template DNA residues in SARS-CoV-2 mRNA vaccine demonstrates high sensitivity, excellent stability, strong specificity, and high accuracy, making it a suitable approach for the detection of template DNA residues in SARS-CoV-2mRNA vaccine.

    2025 04 v.38 [Abstract][OnlineView][Download 977K]

  • Development and verification of ion-pairing RP-HPLC method for determination of residual L-methionine sulphoximine in recombinant protein therapeutics

    WANG Lizhi;YANG Jiaming;LI Benqiang;LI Xiaocui;LIANG Qiuyi;DENG Qinpei;Zhuhai Livzon Mabpharm Inc.;

    Objective To develop an ion-pairing reversed-phase high performance liquid chromatography(IP-RP-HPLC)method for the determination of L-methionine sulphoximine(MSX) residues in recombinant protein therapeutics, and to verify and preliminarily apply it, so as to provide a reference for the quality control of related products.Methods A chromatographic analysis method was screened and the mobile phase conditions such as ion-pairing reagent, pH and salt concentration were optimized to develop an IP-RP-HPLC method for the determination of MSX residues in recombinant protein therapeutics. The method was verified for specificity, system applicability, linearity, limit of detection(LOD) and limit of quantitation(LOQ), precision, accuracy and solution stability, by which the residual MSX contents in multiple batches of in-process sample and bulk of recombinant protein therapeutics were determined.Results The chromatographic conditions were determined as follows: using Hypersil GOLDTMC18 column(4. 6 mm × 250 mm, 5 μm) at column temperature of 25 ℃; 50 mmol/L sodium dihydrogen phosphate(containing 5 mmol/L sodium 1-octane sulfonate, pH 2. 0)-acetonitrile(volume fraction of 85 ∶ 15) as mobile phase for isocratic elution at a flow rate of 0. 6 mL/min; detection wavelength of 200 nm; injection volume of 50 μL. The method exhibited good specificity and system applicability. The control substance showed good linearity in the concentration range of 10-100 μg/mL, with a correlation coefficient(R) of 0. 999 95. The LOD and LOQ were 5 and 0. 5 μg/mL, respectively. MSX was not detected in either recombinant protein A or B, and the relative standard deviations(RSDs) of the measured concentrations of six parallel spiked samples were 1. 3% and 1. 1%, respectively, with the recovery rates ranged from 94% to 103%. For recombinant protein A spiked test sample placed at 10 ℃, the RSD of the measured concentrations at 0 h and 12 h were 2. 1%. MSX was not detected in the in-process samples and bulk of recombinant protein A test samples and three batches of recombinant protein B samples.Conclusion The developed IP-RP-HPLC method has the characteristics of simple operation and high sensitivity, and can be used for the quality control of MSX residues in recombinant protein therapeutics.

    2025 04 v.38 [Abstract][OnlineView][Download 908K]

  • Establishment and verification of gas chromatography method for determination of 2-phenoxyethanol in Sabin inactivated poliovirus vaccine(Vero cells)

    CHENG Bin;HE Wenzhi;JIANG Yiheng;WANG Li;Ministry of Industry and Information Technology Industrial Technology Basic Public Service Platform,Yunnan Institute for Food and Drug Control;

    Objective To establish and verify a gas chromatography(GC) method for the determination of 2-phenoxyethanol content in Sabin inactivated poliovirus vaccine(sIPV)(Vero cells), thereby providing a reliable approach for detecting 2-phenoxyethanol content in biological products. Methods A polyethylene glycol(PEG) GC column(30 m × 0. 25 mm ×0. 25 μm) was employed for the determination of 2-phenoxyethanol content. The chromatographic conditions were as follows:nitrogen as the carrier gas, an injection volume of 1 μL, a split ratio of 1∶ 10, and a carrier gas flow rate of 1 mL/min.The temperature program was set with an initial column temperature of 90 ℃, followed by a ramp at a rate of 10 ℃ per minute to 220 ℃, which was maintained for 10 minutes. The signal was collected by flame ionization detector(FID). The method was verified for the linear range, repeatability, accuracy, and specificity. The established GC method and high-performance liquid chromatography(HPLC) were used to analyze 2-phenoxyethanol content in three batches of sIPV(Vero cells)respectively, and the results were compared. Results The reference solution exhibited a good linear relationship with the peak area in the concentration range of 0. 2-1. 0 mg/mL, with the linear equation: y = 4 021. 44 x + 97. 07, R2= 0. 998. The relative standard deviation(RSD) of 2-phenoxyethanol content in six test solutions was less than 2. 0%. The spiked recovery rates of high, medium, and low concentrations of 2-phenoxyethanol in nine test samples ranged from 90% to 110%. Both the reference and test solutions showed a distinct chromatographic peak for 2-phenoxyethanol at 9. 1 min, while no such peak was observed in the negative control, indicating no interference with the test samples. The mean 2-phenoxyethanol content in three batches of test samples, as determined by GC and HPLC, was(4. 990 ± 0. 175) and(4. 986 ± 0. 122) mg/mL, respectively, with no statistically significant difference(t = 0. 045 6, P = 0. 967 8). Conclusion The established GC method demonstrates good repeatability, accuracy, and specificity, showing potential as a viable alternative to HPLC for the quantification of 2-phenoxyethanol.

    2025 04 v.38 [Abstract][OnlineView][Download 867K]

  • Feasibility study of process scale-up for production of human prothrombin complex concentrate

    ZHAO Haiping;YANG Yu;XIA Yunkong;ZHAO Lei;GAO Zhiqiang;Chengdu Rongsheng Pharmaceuticals Co., Ltd.;

    Objective To evaluate the feasibility of process scale-up for the production of human prothrombin complex concentrate(PCC) by comparing the quality parameters of PCC samples obtained at different stages of process scale-up.Methods The PCC production process was scaled up sequentially through bench scale, pilot scale and production scale.Samples were collected at critical process control points across the three scales for comparative quality analysis. The final PCC products from each scale were tested in accordance with the Chinese Pharmacopoeia(VolumeⅢ, 2020 edition) to assess process stability and regulatory compliance during scale-up.Results After the first ultrafiltration step, no statistically significant differences were observed in the potency of human coagulation factor Ⅸ(FⅨ) or protein content among samples from the three scales(F = 1. 066 and 0. 590, respectively, each P > 0. 05). The FⅨ recovery rates were(69. 3 ± 10. 3)%,(73. 9 ±11. 1)%, and(69. 8 ± 7. 3)%, respectively, with no significant difference( F = 0. 330, P > 0. 05). Following solvent/detergent(S/D) treatment, the pH remained stable, and no significant differences were observed in FⅨ potency or protein content(F =1. 414 and 0. 542, respectively, each P > 0. 05). After the secondary ion-exchange chromatography step, no significant differences were found in FⅨ potency or specific activity(F = 0. 437 and 0. 201, respectively, each P > 0. 05), with FⅨ recovery rates of(90. 6 ± 6. 7)%,(82. 6 ± 4. 6)% and(87. 2 ± 6. 1)%, respectively, with no significant difference(F = 2. 513, P > 0. 05).At the bulk solution stage, no significant differences were observed in FⅨ potency or specific activity(F = 0. 187 and 0. 135,respectively, each P > 0. 05) with stable pH, and FⅨ recovery rates were(90. 6 ± 7. 5)%,(97. 2 ± 8. 3)%, and(92. 2 ± 6. 4)%,respectively, with no significant difference(F = 1. 016, P > 0. 05). After dry-heat virus inactivation, no significant differences were noted in the potency of factorsⅡ, Ⅶ, Ⅸ, and Ⅹ(F = 0. 11, 0. 473, 0. 818, and 0. 244, respectively, each P > 0. 05).The critical quality attributes of final PCC products from all three scales were consistent and complied with the requirements of Chinese Pharmacopoeia(Volume Ⅲ, 2020 edition).Conclusion The established PCC production process is stable, reliable, and reproducible, demonstrating the feasibility of process scale-up.

    2025 04 v.38 [Abstract][OnlineView][Download 862K]

  • Development and verification of RP-HPLC for determination of benzalkonium bromide content in human interferon α1b spray

    LIU Linlin;ZHANG Tinghui;SUN Ruixin;SU Chang;LIU Bingying;LIU Jinghui;The Second Laboratory, Changchun Institute of Biological Products Co., Ltd.;

    Objective To develop a reversed-phase high performance liquid chromatography(RP-HPLC) method for the determination of benzalkonium bromide content in human interferon α1b(hIFNα1b) spray, and to verify and preliminarily apply the method in order to provide a reference for the dosage control of benzalkonium bromide in hIFNα1b spray.Methods The conditions of liquid chromatography were as follows: ZORBAX 300SB-C18 column(4. 6 mm × 250 mm, 5 μm); mobile phase was acetonitrile and 5 mmol/L ammonium acetate solution at a volume ratio of 65∶35; detection time was 30 min at flow rate of 1. 0 mL/min and column temperature of room temperature; UV detection wavelength was 262 nm; injection volume was 20 μL; isocratic elution. The system adaptability, specificity, linearity, accuracy, precision, limit of quantitation(LOQ),limit of detection(LOD), and robustness of the method were verified, and the content of benzalkonium bromide in three batches of hIFNα1b spray was determined by using this method.Results The method demonstrated good system applicability and specificity for benzalkonium bromide detection. A good linear relationship was observed within the concentration range of 10-100 μg/mL(R2 = 0. 999 7). The overall mean spiked recovery rate of accuracy verification was 97. 2% with an RSD of1. 61%. The RSDs of reproducibility and intermediate precision verification were 3. 36% and 1. 20%, respectively. The LOQ and LOD were 0. 02 and 0. 008 μg/mL, respectively. Minor variations in flow rate, mobile phase pH, and column temperature within specified ranges showed no significant impact on the analytical results. Quantitative analysis of three batches of hIFNα1b spray showed benzalkonium bromide contents of 45. 4, 45. 0, and 45. 2 μg/mL, respectively, all conforming to the labeled specification range of 40-60 μg/mL.Conclusion The RP-HPLC method has good specificity, accuracy, precision and robustness, and can be used for the determination of benzalkonium bromide in hIFNα1b spray.

    2025 04 v.38 [Abstract][OnlineView][Download 901K]

  • Research progress and application prospects of circular RNA

    HUANG Tianzhi;SHI Xinchang;ZHOU Yong;Division of Recombinant Biological Products, National Institutes for Food and Drug Control,Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech Products;

    Circular RNA(circRNA) is a type of RNA with a circular coding structure, characterized by a circular topological structure formed during the RNA splicing process. Compared to traditional linear mRNA, circRNA exhibits enhanced stability and immunogenicity, leading to its wide-ranging potential applications in areas such as vaccines, biomarkers, and targeted therapies. This paper summarizes the research progress of circRNA, including its synthesis and degradation in vivo and in vitro, components, biological functions, and applications, aiming to provide novel insights for their utilization in vaccine design and development.

    2025 04 v.38 [Abstract][OnlineView][Download 1025K]

  • Advances in vaccines against urinary tract infection

    YU Cheng;HE Runming;NIU Hongxia;College of Basic Medical Science, Zhejiang Chinese Medical University;

    Urinary tract infections(UTIs) are prevalent disorders of the urinary system, primarily caused by pathogens such as uropathogenic Escherichia coli(UPEC). These infections significantly impair patients' quality of life and increase the risk of urosepsis. Although antibiotic therapy can alleviate symptoms, prolonged use often leads to drug resistance and recurrence, underscoring the necessity for developing novel prevention and treatment strategies for UTIs. In recent years, UTI vaccines, as a specific preventive approach, have shown promising progress, which are broadly categorized into three types:whole-cell/lysate vaccines, live attenuated vaccines, and subunit vaccines. Whole-cell/lysate vaccines, such as Uromune,Strovac, and OM-89/UroVaxom, have already been commercialized. In contrast, live attenuated vaccines and subunit vaccines, including ExPEC4V and ExPEC10V, remain in the preclinical research stage. In this paper, the mechanism underlying UTIs and the recent advancements in UTI vaccine development are reviewed, aiming to provide a reference for the research and development of UTI vaccines in China.

    2025 04 v.38 [Abstract][OnlineView][Download 934K]

  • Research progress on effect of lactoferrin on tissue injury repair

    ZHAO Yu;ZHANG Bowen ;SHU Xingfu;ZHANG Haixia;Key Laboratory of Biotechnology and Bioengineering of State Ethnic Affairs Commission,Biomedical Research Center, Northwest Minzu University;

    Lactoferrin(LF) belongs to the transferrin family and is a multifunctional glycoprotein with a variety of biological activities, which not only binds to viral antigenic determinants to inhibit the recognition and binding of viruses to host cells,but also inhibits cell carcinogenesis through the production of thiobarbituric acid, malondialdehyde and other substances with strong antioxidant properties. In recent years, it has been found that in addition to its antiviral and anti-inflammatory functions, LF also has the ability to promote the repair of various types of damaged tissues. This paper mainly reviews the repairing and pro-repairing effects of LF after skin, muscle, bone, intestinal, brain and gingival injuries, aiming to elucidate the further application prospects of LF in the medical field.

    2025 04 v.38 [Abstract][OnlineView][Download 844K]

  • Advances in anti-tumor effects of macrophage-derived exosomes

    DU Wenya;DAI Yumei;WU Lixian;Department of Microbiology and Immunology, School of Basic Medical Sciences,Dali University;

    Tumors are highly heterogeneous diseases, and tumor-associated macrophages(TAMs) are important immune cells in the tumor microenvironment, which are highly heterogeneous and plastic. Tumor recurrence and metastasis are an important cause of morbidity and mortality in cancer patients. Macrophage-derived exosomes(MDEs) act as messengers for cell-to-cell signaling and carry a variety of components of mother cells. A large number of studies have shown that the polarization state of macrophages plays a regulatory role in tumor progression, and MDE is closely related to tumor progression and macrophage polarization. As a new anti-tumor strategy, MDE provides new insights into tumor treatment. This paper summarizes recent research on engineered MDEs in anti-tumor and how M1 and M2 MDEs mediate tumor development, aiming to provide references for tumor treatment.

    2025 04 v.38 [Abstract][OnlineView][Download 879K]

  • Effect of exosome-mediated intercellular communication on invasion and metastasis of hepatocellular carcinoma

    LIU Qianqian;SUN Xinglu;LIU Fang;CHEN Che;School of Public Health, Gansu University of Chinese Medicine;

    Hepatocellular carcinoma(HCC) is the most common primary liver malignancy with high metastasis and recurrence rates, and is one of the leading causes of cancer-related deaths worldwide. Despite improvements in detection and treatment, the prognosis for HCC remains poor. In recent years, exosomes, as a newly discovered intercellular communication mediator in the occurrence and development of tumors, deliver biologically active substances to activate or inhibit various signal transduction pathways in recipient cells, thereby regulating liver cancer cells including proliferation, invasion,metastasis, drug resistance and other malignant behaviors. However, the mechanism of exosomes on the invasion and metastasis of HCC is still unclear. Therefore, this paper systematically reviewed the regulatory mechanism of exosomes in HCC invasion and metastasis.

    2025 04 v.38 [Abstract][OnlineView][Download 868K]

  • Research progress on epidemiological characteristics and prevention of herpes zoster in China

    ZHOU Hui;SHUANG Hui;ZHANG Fukun;Changchun Keygen Biological Products Co., Ltd.;

    Herpes zoster(HZ) is an acute infectious skin disease caused by the reactivation of varicella-zoster virus(VZV)which remains latent in the posterior root ganglion of spinal cord or cranial ganglion for a long time. It usually causes skin lesions such as erythema and herpes, accompanied by different degrees of pain or nerve-related symptoms. HZ mostly occurs in middle-aged and elderly people, as well as people with low immunity. The aging population in China is becoming increasingly severe, and the disease burden and public health problems brought about by it have gradually become prominent. In this paper, the epidemiological characteristics and preventive measures of HZ in China are reviewed, aiming to provide references for the prevention and control of HZ in China.

    2025 04 v.38 [Abstract][OnlineView][Download 863K]
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@2012 Editorial Department of "Chinese Journal of Biologicals"