Current Issue

  • Preparation and immunogenicity evaluation of a tick-borne encephalitis mRNA vaccine producing virus-like particles

    LI Zhuang;GUO Yibing;ZHAO Chen;ZHOU Hang;WU Yehong;Vaccine Research and Development Department, Changchun Institute of Biological Products Co., Ltd.;

    Objective To prepare an mRNA vaccine against tick-borne encephalitis virus(TBEV), verify its ability to produce virus-like particles(VLPs) in cells, and evaluate its immunogenicity in mice.Methods The gene sequence containing Japanese encephalitis virus(JEV) signal peptide, TBEV premature membrane glycoprotein(prM) and E protein was designed. The plasmids were constructed according to the sequence, and corresponding mRNA was synthesized by in-vitro transcription. The mRNA was transfected into HEK293T cells and then measured for the expression in cells by Western blot,and detected for the ability to self-assemble into VLPs in cells by electron microscopy. The mRNA lipid nanoparticles(LNPs)were prepared by Nano Assembler system, and the particle size and distribution of LNPs were measured. Female BALB/c mice were immunized by intramuscular injection at dosages of 2 and 10 μg mRNA vaccine respectively, 6 mice in each group, and the negative control group(PBS) was set. One week after booster immunization, orbital blood samples were collected and separated for serum. The binding antibody levels were detected by ELISA, and the neutralizing antibody levels were detected by plaque reduction assay.Results After mRNA transfection into HEK293T cells, a single band was found at the relative molecular mass of about 55 000, which was consistent with the theoretical relative molecular mass of E protein.Under electron microscope, spherical particles with a diameter of about 50 nm were observed, and there were obvious spikes on the surface. The average particle size of mRNA LNPs vaccine was 124. 4 nm and the polymer dispersity index(PDI) was0. 049. Compared with the negative control group, the levels of binding antibody and neutralizing antibody induced by 2 and10 μg mRNA significantly increased(t = 12. 4 and 10. 8, 19. 7 and 12. 0, respectively, each P < 0. 001).Conclusion A TBEV mRNA vaccine capable of producing VLPs in cells was successfully prepared, which demonstrated good immunogenicity in mice, providing experimental basis for the development of new generation TBEV vaccine in combination using of VLPs technology and mRNA technology.

    2025 08 v.38 [Abstract][OnlineView][Download 905K]

  • Application of trypsin digested-polypeptone medium in preparation of diphtheria vaccine adsorbed

    WEN Jiana;GU Qin;LI Jingyan;PING Ling;ZHANG Yunkun;YU Min;DU Zhiyun;YANG Jingsi;Institute of Medical Biology of Chinese Academy of Medical Science & Peking Union Medical College, Kunming Science and Technology Innovation Center for the Research and Development of Vaccines against New Outbreaks of Highly Pathogenic Pathogens;

    Objective To explore the possibility of using trypsin digested-polypeptone medium to prepare diphtheria vaccine adsorbed, so as to improve the quality and safety of the vaccine.Methods Corynebacterium diphtheriae(C.diphtheriae) was cultured in trypsin digested-polypeptone medium(polypeptone group) and modified Linggood's medium(modified Linggood's group), and three batches were cultured. The culture solution of C.diphtheriae was collected, and the toxin production detection, SDS-PAGE analysis and viable bacteria count were performed. Diphtherotoxin(DT) was extracted, and the 50% tissue culture infective dose(TCID50) was measured, and the logarithm was taken. After formaldehyde detoxification treatment, DT was tested for purity and analyzed by SDS-PAGE. Diphtheria vaccine adsorbed was prepared according to the corresponding method of Chinese Pharmacopoeia(VolumeⅢ, 2020 edition), and the vaccine titer was detected.Results The mean toxin production of three batches of culture solution of C.diphtheriae in polypeptone group was 177 Lf/m L, slightly lower than that of modified Linggood′s group(183 Lf/m L), and there was no statistically significant difference between them(t = 2. 00, P > 0. 05).The relative molecular weights of toxin protein and diphtheria toxoid obtained from two kinds of culture media were about 60 000. The mean values of A600and viable bacteria, lg TCID50of DT, purity of diphtheria toxoid and titer of diphtheria vaccine adsorbed in modified Linggood's group were significantly higher than those in polypeptone group(t = 9. 390, 31. 180,4. 914, 44. 540 and 4. 510, respectively, each P < 0. 05), while all the indicators in polypeptone group met the relevant requirements of Chinese Pharmacopoeia(VolumeⅢ, 2020 edition). Conclusion Trypsin digested-polypeptone medium is expected to replace the traditional modified Linggood's medium for the preparation of diphtheria vaccine adsorbed, while the preparation technology needs to be further optimized to improve the vaccine quality.

    2025 08 v.38 [Abstract][OnlineView][Download 893K]

  • Evaluation of protective effect of two virus protective agents against a novel varicella-herpes zoster virus(Oka-7S)

    LI Hang;XU Junfeng;JIANG Xueou;WEI Bo;LI Meng;FANG Jia;ZHANG Fukun;Research Office, Changchun Keygen Biological Products Co., Ltd.;

    Objective To compare the effects of freeze-thaw, ultrasound and temperature on the novel varicella-herpes zoster virus(VZV)(Oka-7S) under different virus protective agents.Methods Oka-7S virus samples(intracellular and extracellular virus samples) were freeze-thawed for four times under No.1 and No.2 virus protective agents at-80 ℃, and the virus titer was detected at each freeze-thaw. Oka-7S virus samples(intracellular and extracellular virus samples) under different virus protective agents were ultrasonically studied at 20 kHZ for six times, and detected for the virus titer after each ultrasound. The survival time of Oka-7S virus samples(intracellular and extracellular virus samples) under different virus protective agents was detected at 4 ℃, 25 ℃ and 37 ℃.Results Protective agent No.2 had the best protective effect on Oka-7S intracellular and extracellular virus samples. The titer of Oka-7S intracellular virus samples was higher at-80 ℃, and the effect of freeze-thaw once was the best. Under ultrasonic conditions, protective agent No.2 made Oka-7S intracellular virus release more virus particles, and at the same time, Oka-7S extracellular virus samples had higher titer. The Oka-7S intracellular virus samples had the best effect at 20 kHZ for 3 s, while the Oka-7S extracellular virus samples had the best effect at20 kHZ for 1 s. In addition, the survival time of Oka-7S intracellular and extracellular virus samples was longer at 4 ℃, and the preservation time of Oka-7S virus samples was significantly prolonged by protective agent No.2 compared with protective agent No.1.Conclusion Under different preservation conditions, the protective effect of No.2 protective agent on virus was better under freeze-thaw, ultrasound and temperature conditions, and the titer loss of intracellular and extracellular virus samples of Oka-7S was lower.

    2025 08 v.38 [Abstract][OnlineView][Download 972K]

  • Effect of corticosterone on expression of brain-derived neurotrophic factors in CA1 region of rat hippocampus and its mechanism

    LIU Jialin;DAI Caixing;JIANG Zhihua;ZHANG Xinyue;YAN Zi;ZHAO Xin;LI Jianguo;Key Laboratory of Cellular Physiology, Ministry of Education, Department of Physiology,Shanxi Medical University;

    Objective To investigate the effect of corticosterone(CORT) on the expression of brain-derived neurotrophic factors(BDNFs) in the CA1 region of rat hippocampus and its mechanism, so as to provide new ideas for the study of the pathogenesis of depression.Methods A rat model with depressive-like behavior was established by long-term oral administration of CORT, and the control group was set up(no administration). Western blot was used to detect the expression levels of BDNFs and precursor BDNF(proBDNF) in the CA1 region of rat hippocampus. In situ hybridization(ISH) assay was used to detect the expression distribution of Bdnf transcripts Ⅰ、Ⅱ、Ⅳ and Ⅵ in the CA1 region of the hippocampus. Chromatin immunoprecipitation(ChIP) assay was used to detect the trimethylation of histones H3K4, H3K9, H3K27 and H3K36 in the promoters p1, p2, p4 and p6 of Bdnf gene in the CA1 region.Results Compared with the control group, the expression of BDNF protein in the CA1 region of rat hippocampus in the CORT group significantly decreased(t = 2. 842, P < 0. 05),while the expression of proBDNF protein significantly increased(t = 3. 227, P < 0. 05); the expression of Bdnf transcripts Ⅰ,Ⅱ,Ⅳ and Ⅵ decreased; the trimethylation levels of histones H3K4, H3K9, H3K27 and H3K36 in the p1 region of Bdnf promoter significantly increased(t = 17. 280 0, 4. 649 0, 10. 480 0, and 46. 570 0, respectively, each P < 0. 001); there was no significant difference in the trimethylation levels of the four sites in p2 and p4 regions of Bdnf promoter(t = 0. 105 1-2. 633 0, each P > 0. 05); the trimethylation level at H3K27 in the p6 region significantly increased(t = 5. 383 0, P < 0. 018), while there was no significant difference at H3K4, H3K9 and H3K36(t = 2. 022 0, 0. 875 0, 0. 486 2, respectively, each P > 0. 05).Conclusion Oral administration of CORT can induce depression-like behaviors in rats, and cause the significantly decreased expression of BDNF protein and significantly increased expression of proBDNF protein in the CA1 region of rat hippocampus in CORT group, which may be related to the decreased expression of Bdnf transcripts Ⅰ、Ⅱ、Ⅳ and Ⅵ, as well as different histone methylation modification patterns in the p1, p2, p4 and p6 regions of Bdnf promoter.

    2025 08 v.38 [Abstract][OnlineView][Download 995K]

  • Expression and identification of Spo11protein in Giardia duodenalis

    WANG Xin;ZHANG Nan ;LI Jianhua;GONG Pengtao;WANG Xiaocen;LI Xin;ZHANG Xu;ZHANG Xichen;College of Veterinary Medicine, Jilin University;

    Objective To express and identify Spo11 protein of Giardia duodenalis(Giardia)using prokaryotic and eukaryotic expression system.Methods The Spo11 gene was amplified by PCR, then recombined into pGEX-4T-1 prokaryotic vector and transformed into competent E.coli DH5α. After sequencing, the positive plasmid was transformed into E.coli BL21(DE3)and induced by IPTG. The obtained Spo11 protein was purified by GST Resin affinity chromatography and identified by Western blot. The Spo11 gene was recombined into the pcDNA3.1 eukaryotic expression vector and transformed into the competent E.coli DH5α. After sequencing, the endotoxin-free plasmid was extracted and transfected into HEK293T cells, and the cells were collected and lysed for Western blot analysis.Results The amplified fragment of Spo11gene of Giardia was 1 107 bp in length, and the recombinant plasmid pGEX-4T-1-Spo11 was constructed correctly as identified by double enzyme digestion and sequencing. The purified GST-Spo11 protein, with a relative molecular mass of about 50 000, showed specific binding to GST-tag antibody and good reactivity. The plasmid pcDNA3.1-HA-Spo11 was constructed correctly as identified by double digestion and sequencing. The expressed HA-Spo11 protein had a relative molecular mass of about 40 000, which showed specific binding to HA-tag antibody with good reactivity.Conclusion The Spo11 protein was successfully expressed by prokaryotic and eukaryotic expression system, which laid a foundation for the study of Spo11 protein function in Giardia.

    2025 08 v.38 [Abstract][OnlineView][Download 1028K]

  • Analysis of metabolites of Semaglutide in human liver microsomes

    LI Ju;LIN Tao ;YANG Lei ;LI Yan ;ZHOU Chao;Deyang Hospital Affiliated Hospital of Chengdu University of Traditional Chinese Medicine;

    Objective To analyze the metabolic characteristics and major metabolites of Semaglutide, a glucagon-like peptide-1(GLP-1) receptor agonist, in human liver microsomes.Methods Human liver microsomal incubation system was used to metabolize Semaglutide in vitro. The metabolites were identified by ultra-performance liquid chromatography(UPLC) and X500R quadrupole time-of-flight(QTOF) mass spectrometry.Results Semaglutide underwent extensive metabolism in human liver microsomes, mainly through proteolytic cleavage of peptide skeleton and oxidation of amino acid residues. A total of six metabolites were identified, five of which were discovered for the first time.Conclusion The discovery of Semaglutide metabolites in human liver microsomes provides important information for further study of the pharmacodynamic properties and safety, and contributes to a deeper understanding of the metabolite characteristics of Semaglutide and its implications for clinical application.

    2025 08 v.38 [Abstract][OnlineView][Download 1165K]

  • Expression and clinical significance of GINS2 in endometrial cancer tissues

    LUO Jianbo;PAN Yuqing ;ZHAI Li;TAO Pengzuo;ZHU Yingfang;FENG Zhiqi;ZHANG Xi;Department of Clinical Laboratory, Yunnan Cancer Hospital, The Third Affiliated Hospital of Kunming Medical University;

    Objective To investigate the expression of GINS2 in endometrial cancer(EC) tissues and its correlation with prognosis.Methods The differential expression of GINS2 gene in EC and normal control tissues was analyzed using GEPIA,TIMER, and UALCAN databases. The endometrial tissues of 115 patients with EC(EC group), 30 patients with uterine fibroid(uterine fibroid group) and 26 patients with normal endometrium(normal control group) treated and followed up from May 2017 to December 2022 at the Department of Gynecology, Yunnan Cancer Hospital were collected. The transcription and translation levels of GINS2 gene in the samples were detected by qPCR and Western blot, respectively. Additionally, the expression levels and localization of GINS2 in different samples were analyzed using immunohistochemistry. Furthermore,the correlation between GINS2 expression levels and the clinicopathological features of EC was analyzed, and potential independent prognostic factors for EC were evaluated using Kaplan-Meier survival analysis and Cox proportional hazards regression model.Results The mRNA expression level of GINS2 gene was significantly higher in EC tissues than in normal control tissues according to all three databases(P < 0. 05). In the clinical validation cohort, both the transcriptional and protein expression levels of GINS2 gene were significantly higher in the EC group than in the uterine fibroid group and normal control group(t = 7. 707 and 8. 330, respectively, each P < 0. 01). GINS2 protein was mainly located in the cell nucleus and partially in the cytoplasm of EC tissues, with positive expression rates of 81. 74%, 46. 67% and 38. 46% in EC tissues,uterine fibroids, and normal endometrial tissues, respectively. GINS2 levels in EC tissues were associated with differentiation degree and lymph node metastasis(t = 4. 125 and 3. 589, respectively, each P < 0. 05), but not with age, tissue type, International Federation of Gynecology and Obstetrics(FIGO) stage, depth of invasion, estrogen receptor(ER) expression, or progesterone receptor(PR) expression(t = 0. 214, 0. 247, 0. 125, 0. 211, 0. 238, and 0. 314, respectively, each P > 0. 05). Multivariate Cox regression analysis showed that elevated GINS2 expression [hazard ratio(HR) = 2. 091, 95% confidence interval(CI): 1. 048-3. 135, P = 0. 019] and lymph node metastasis(HR = 1. 106, 95% CI: 1. 064-1. 149, P = 0. 027) were independent prognostic factors for EC patients.Conclusion The high expression of GINS2 in tissues is associated with poor prognosis of EC patients, and it is speculated that it may be related to the occurrence and development of EC.

    2025 08 v.38 [Abstract][OnlineView][Download 1198K]

  • Surveillance of measles, rubella and mumps antibody levels in healthy population in Pudong New Area of Shanghai from 2018 to 2022

    YANG Dandan;LIU Wenmin;WANG Weiping;YANG Laibao;DENG Pengfei;XUE Caoyi;FEI Yi;Department of Immunology and Prevention, Shanghai Pudong New Area Center for Disease Control and Prevention(Shanghai Pudong New Area Health Supervision Institute);

    Objective To monitor the antibody levels against measles, mumps and rubella in healthy population in Pudong New Area of Shanghai from 2018 to 2022, so as to provide evidence for the improvement of immunization strategy.Methods A total of 1 305 healthy people from 16 communities in Pudong New Area, Shanghai were randomly sampled and divided into12 age groups(0 ～ < 8 months old to ≥ 50 years old). The levels of IgG antibodies against measles, rubella and mumps in venous blood serum were detected by ELISAResults One sample was discarded because no results were detected. In 1 304serum samples, the positive rates of measles, rubella and mumps IgG antibodies were 63. 65%, 48. 39% and 58. 21%, and the geometric mean concentrations(GMCs) of the antibodies were 229. 74 mIU/mL, 11. 63 IU/mL and 76. 43 U/mL, respectively.The positive rates of measles, rubella and mumps IgG antibodies(χ~2= 0. 01-4. 874, each P > 0. 05) and GMCs(t = 0. 020-2. 583,P > 0. 05) in the serum of healthy people of different genders and household registrations showed no statistically significant difference, while the positive rates of IgG antibodies(χ~2= 16. 865-667. 297, each P < 0. 01) and GMCs(t = 5. 862-117. 019,each P < 0. 001) in the serum of healthy people with different age, year and vaccination history exhibited statistically significant differences.Conclusion The antibody levels of measles, rubella and mumps among healthy population in Pudong New Area of Shanghai are related to the year, age and immunization history, and there is still a certain risk of infection. It is suggested to strengthen the monitoring of immune status of healthy population and vaccination of women at childbearing age related to measles, mumps and rubella, which will help reduce the risk of measles, rubella and mumps in healthy population under vaccination age.

    2025 08 v.38 [Abstract][OnlineView][Download 900K]

  • Identification of different variant strains of inactivated SARS-CoV-2 vaccines(Vero cells) based on UPLC-Q-Orbitrap-MS/MS analysis technology

    YAN Ping;GUO Jing ;YANG Anna ;XIANG Meifeng ;SHANG Jun ;HE Kaiyong;Hubei Institute for Drug Control, NMPA Key Laboratory of Quality Control of Blood Products;

    Objective Toestablishanultra-performanceliquidchromatography-quadrupole-electrostaticfieldorbitraptandem mass spectrometry(UPLC-Q-Orbitrap-MS/MS) analysis technology for the identification of different variants of inactivated SARS-CoV-2 vaccines(Vero cells), so as to provide a new evaluation direction for vaccine supervision.Methods Peptide samples were obtained from inactivated SARS-CoV-2 vaccines after reductive alkylation, tryptic digestion, and desalting of SDB-RPS desalting column, and then were eluted by CDS C18 column(0. 075 mm × 250 mm × 1. 9 μm) in gradient program, with 0. 1% formic acid and 3% dimethyl sulfoxide as mobile phase A, 0. 1% formic acid, 3% dimethyl sulfoxide and 80% acetonitrile as mobile phase B. The column temperature was 50 ℃, the flow rate was 300 nL/min, and the injection volume was 5 μL. Then UPLC-Q-Orbitrap-MS/MS analysis technology was established, the mass spectrometry detection was scanned by data-dependent acquisition(DDA) mode, and the sample peptide information was retrieved and analyzed by MaxQuant(V 2.4.2.0) software.Results The established method was used to analyze and determine the inactivated SARSCoV-2 vaccines(Vero cells) of different variants, and the conserved peptides and characteristic peptide sequences of S protein and N protein of different variants of SARS-CoV-2 were screened and extracted, and the peptide mass-to-charge ratio(m/z) was combined to obtain the specific protein database of different variants. By comparing the peptide information detected by the mass spectrometry of the sample, it is possible to identify different variants of inactivated SARS-CoV-2vaccines(Vero cells).Conclusion The UPLC-Q-Orbitrap-MS/MS analysis technology established in this study has the advantages of high sensitivity, rapidity and strong specificity, which can be effectively used for the identification and screening of strains of inactivated SARS-CoV-2 vaccines(Vero cells), can provide a technical reference for the identification of SARS-CoV-2 species in other viral vaccines or biological samples, and provide a new perspective for the quality and safety supervision of vaccines.

    2025 08 v.38 [Abstract][OnlineView][Download 942K]

  • Application of mixed-mode chromatography in development of bispecific antibody purification process

    WEI Li;LIU Daoqin;YU Xuelian;XU Ting;CHEN Chuchu;GAO Hailing;WANG Chunyan;WU Tingting;WANG Ruyi;DOU Yinghui;FAN Qinglin;SONG Lihua;Anhui Anke Biotechnology (Group) Co., Ltd.;

    Objective To eliminate the aggregates during the production process of bispecific antibodies(BsAbs) by employing the mixed-mode chromatography, and to enhance product purity and eradicate residual impurities while simultaneously ensuring optimal recovery rates of target proteins.Methods The purification process of an IgG-like BsAb in flowthrough mode was developed using a mixed-mode chromatography(anionic and hydrophobic) combined with the Design of Experiments(DoE). A purification process with high purity and high recovery rate was established. The critical process parameters of mixed-mode chromatography in flow-through mode were identified, including the pH and conductivity of sample loading solution and equilibration buffer, retention time, sample loading capacity, as well as their respective effects on aggregate removal and the distribution of aggregates at different peak fractions.Results After implementing a three-step chromatographic purification procedure on the CHO cell culture supernatant containing BsAb, the size exclusion highperformance liquid chromatography(SEC-HPLC) analysis revealed that the purity of the target BsAb product exceeded99%, with residual host cell proteins(HCPs) at levels below 20 ppm and undetectable levels of residual host cell DNA. ConclusionMixed-mode chromatography exhibits a broad range of potential applications in the development of Bs Ab purification processes, as it effectively eliminated approximately 7% aggregates while maintaining the recovery exceeding 93% for the target protein in this study.

    2025 08 v.38 [Abstract][OnlineView][Download 1151K]

  • Development and verification of HPLC for determination of residual EDTA disodium salt in freeze-dried rabies vaccine(Vero cells) for human use

    FENG Ziyi;LI Yanxia;LIN Hongjuan;XIONG Jinfeng;WEI Dongdong;LI Chunyan;SUN Hongliang;Changchun Institute of Biological Products Co., Ltd.;

    Objective To develop a high-performance liquid chromatography(HPLC) method for the determination of residual EDTA disodium salt(EDTA-2Na) content in freeze-dried rabies vaccine(Vero cells) for human use, and to verify and preliminarily apply the method, so as to ensure the content of EDTA-2Na within the safe concentration range.Methods The condition for HPLC was as follows: reverse chromatographic column was adopted, using the mobile phase consisting of 0. 1%phosphate solution as the aqueous phase and 100% methanol as the organic phase at a flow rate of 1. 0 mL/min and a detection wavelength of 242 nm. The injection volume was 20 μL, and the column temperature was 25 ℃. The developed method was verified for suitability, limit of quantitation(LOQ), specificity, linear range, accuracy, reproducibility, and intermediate precision, and the residual EDTA-2Na contents in thr ee batches of bulk of freeze-dried rabies vaccine(Vero cells) for human use were determined using this method.Results The LOQ of EDTA-2Na detected by this method was 0. 490 0 μg/mL. The blank and sample solution had no interference to EDTA-2Na, and the separation degree of the target and adjacent peaks was more than 1. 5. EDTA-2Na showed good linearity in the concentration range of 0. 490 0 to 9. 800 0 μg/mL. The recovery rates of EDTA-2Na in spiked samples at concentrations of 50%, 100% and 150% were from 94% to 113%. The recovery rates of EDTA-2Na in six solutions of reproducibility verification ranged from 100% to 102%, with a relative standard deviation(RSD) of 1%. A total of 12 solutions of intermediate precision verification from different personnel were measured, and the RSD of recovery rate was 2%. In addition, EDTA-2Na was not detected in three batches of samples.Conclusion The developed HPLC has good suitability, specificity, linearity, accuracy, reproducibility and intermediate precision, which is suitable for the determination of residual EDTA-2Na in bulk of freeze-dried rabies vaccine(Vero cells) for human use.

    2025 08 v.38 [Abstract][OnlineView][Download 920K]

  • Analysis of N-terminal sequencing results of N-terminal site-specific mono-PEGylated recombinant human granulocyte colony-stimulating factor

    SHI Xinchang;JIAO Xuwen ;WEI Changlong;LIANG Wenyue ;LIANG Weiyang ;LIANG Chenggang;NHC Key Laboratory of Research on Quality and Standardization of Biotech Products; NMPA Key Laboratory for Quality Research and Evaluation of Biological Products, National Institutes for Food and Drug Control;

    Objective To investigate the effect of occupation of modified site of N-terminal site-specific mono-PEGylated recombinant human granulocyte colony-stimulating factor(rh G-CSF)(PEG-G for short) on N-terminal amino acid sequencing based on Edman degradation method.MethodsAccording to the standard sequencing procedure of the N-terminal amino acid sequencer, the N-terminal amino acid sequence of the reference substance for N-terminal site-specific mono-PEGylation of human granulocyte stimulating factor was detected for four cycles, with a sample loading amount of 500 fmol. The waste liquid and mobile phase from the N-terminal amino acid sequencer were collected and evaporated to dryness under low pressure. After dissolving in 2 m L of water, PEG was detected by reversed-phase high performance liquid chromatography(RPHPLC) combined with an evaporative light scattering detector(ELSD).[conditions: mobile phase A, 0. 1% trifluoroacetic acid(TFA)-aqueous solution; mobile phase B, 0. 1% TFA-acetonitrile solution; chromatographic column: C4 column; flow rate: 1. 0 m L/min; injection volume: 100 μL; column temperature: 25 ℃; elution gradient: 0 min→20 min with mobile phase B of 5%→95%, 20 min→30 min with mobile phase B of 95%, 30 min→40 min with mobile phase B of 5%. ELSD conditions: default parameters for carrier gas; temperature 90 ℃; gain 100]. After confirming the PEG retention time, the mobile phase was collected according to the retention time and mass spectrometry analysis was performed.(mass spectrometry collection mode: cation mode; m/z: 3 800-6 000; deconvolution mass tolerance: 20 ppm). N-terminal amino acid sequencing analysis was performed on PEG-G from other companies.ResultsIn the cleaning waste liquid of the N-terminal sequencer, an evaporative light detection chromatographic peak with a retention time close to HPLC was detected. After mass spectrometry analysis, the peak exhibited typical PEG mass spectrometry characteristics, proving that the N-terminal cleaning program(i.e. the first cycle) could cleave the first peptide bond at the N-terminus of some PEG-G, causing the modified PEG to detach. In the sequencing of similar products from other enterprises, it was found that the cracking efficiency was related to the sulfur(S) element near the modified site.ConclusionThe cleaning and sequencing program of N-terminal sequencer can break the N-terminal site-specific PEGylated molecules, which provides technical supports for the establishment of the detection method for N-terminal modified sites in N-terminal site-specific single-modified PEG-G.

    2025 08 v.38 [Abstract][OnlineView][Download 1002K]

  • WHO preferred product characteristics for prevention of respiratory syncytial virus infection in infants and young children

    LI Yufeng;WEN Shifang ;WANG Meixia ;SHI Liwei;Beijing Biological Products Research Association;

    Respiratory syncytial virus(RSV) is one of the main pathogens of lower respiratory tract infections. Although people of all ages can be infected with RSV, the illness tends to be more severe in infants, especially in premature infants,children with chronic lung disease or congenital heart disease, and the elderly, with the majority of the disease burden located in low-and middle-income countries(LMICs). The global availability of safe and effective vaccines is an established principle of the World Health Organization(WHO). Based on this principle, the WHO held two consultations in March 2015and April 2016 to discuss preventive interventions and advance the development of immunoprevention products for RSV infections. In 2017 and 2021, the WHO announced the preferred product characteristics(PPCs) for RSV vaccines and monoclonal antibodies(mAbs). Combined with the relevant guidelines from the WHO and countries with mature research and development, as well as existing market products, this paper comprehensively elaborates on the PPCs recommended by the WHO for immunoprevention products, RSV vaccines and mAbs.

    2025 08 v.38 [Abstract][OnlineView][Download 886K]

  • Research status of production of bioactive peptides by genetic engineering

    YU Tongxin;FANG Yuxin;DONG Na;Animal Science and Technology College, Northeast Agricultural University;

    At present, bioactive peptides have great potential to replace antibiotics as feed additives. It is particularly important to understand the expression system for the production of bioactive peptides and ways to increase their expression. In this paper, the advantages and disadvantages of Escherichia coli, Bacillus subtilis, Pichia pastoris and lactobacillus expression systems were elaborated, the methods to improve the expression of exogenous genes were summarized, and the research progress on the production of bioactive peptides at home and abroad in recent years was also reviewed.

    2025 08 v.38 [Abstract][OnlineView][Download 866K]

  • Research progress on detection methods for hemagglutinin content in influenza vaccines

    ZHENG Aoxue;ZHANG Zhegang;Wuhan Institute of Biological Products Co., Ltd., National Engineering Technology Research Center for Combined Vaccines;

    Influenza is an acute respiratory infection disease caused by influenza viruses, and the main means of preventing influenza is vaccination. Hemagglutinin(HA) is an important component of influenza vaccine that induces the immune response of the body, and accurate and sensitive detection of HA content is of great significance for the development and efficacy evaluation of influenza vaccines. The international gold standard for detecting HA content is single-radial immunodiffusion(SRID), but the production and market time of vaccines are limited due to its dependence on the corresponding standards.In orderto overcome the limitations ofSRID,alternative methods have been explored from two aspects:physicalchemistry and immunochemistry. This paper reviews the research progress on existing methods for detecting HA content in influenza vaccines.

    2025 08 v.38 [Abstract][OnlineView][Download 896K]

  • Research progress on cross-protection of human papillomavirus vaccines

    LIU Huan;GENG Yansheng;HUANG Weijin ;ZHANG Li;Hebei Key Laboratory of Public Health Safety, School of Public Health, Hebei University;

    Human papillomavirus(HPV)is one of the common sexually transmitted viruses in the human reproductive tract,which can cause cervical cancer, genital warts and other related diseases. HPV vaccines could not only prevent infection caused by the same HPV types as vaccines effectively, but also provide additional cross-protection against infection or diseases caused by non-vaccine types. This paper reviewed the cross-protection mechanisms of various types of HPV, the cross-protection effects of the commercial HPV vaccines, as well as the new preventive vaccines in progress, in order to provide a reference for the development of new generation of HPV preventive vaccines with broader protection.

    2025 08 v.38 [Abstract][OnlineView][Download 912K]

  • Research and prospect of coronavirus vaccine technology based on literature and patent quantitative analysis

    SUN Lijing;LIU Xinrui;XIA Huanzhang ;YUAN Hongmei;School of Business Administration, Shenyang Pharmaceutical University;

    The COVID-19 pandemic, which emerged in 2019, presents significant challenges to human health, well-being,and global economic progress. Vaccines are crucial in curbing the spread of viral diseases. This paper provided an overview of scientific research and technological advancements in coronavirus vaccine development, drawing upon literature and patent quantitative analysis approaches to establish a theoretical foundation for future research.

    2025 08 v.38 [Abstract][OnlineView][Download 1199K]

  • Quality by Design-driven systematic development and lifecycle control strategies for polysaccharide conjugate vaccines

    YANG Dan;HU Lin;LI Maoguang ;ZHANG Ying;LI Xiaojing;YE Qiang ;LI Min;Center for Drug Evaluation, National Medical Products Administration;

    The quality of polysaccharide conjugate vaccines is critically dependent on scientific design during early development and lifecycle control. Guided by the Quality by Design(QbD) framework, a systematic development strategy was established, encompassing strain-driven upstream/downstream process development, polysaccharide structure-oriented conjugation process design, carrier protein compatibility optimization, and clinical demand-driven iterative process refinement. By dynamically aligning critical quality attributes(CQAs) with critical process parameters(CPPs), process robustness was ensured. To address complex challenges such as molecular heterogeneity, multi-valent formulation, and batch variability,tailored quality control strategies were proposed based on product characteristics. These encompassed core issues such as functional group-specific control, polysaccharide integrity verification, carrier protein purity assurance, mitigation of crossinterference in multi-valent vaccines, and method selection and cross-validation. Furthermore, the process analytical technology(PAT) and continuous process verification(CPV) were integrated to construct a lifecycle control network spanning early development to commercialization, thereby advancing the alignment of polysaccharide conjugate vaccines with international standards. From a regulatory science perspective, this article systematically elucidates the pivotal role of QbD in safeguarding vaccine quality and clinical efficacy, providing scientific guidance for the rational development, quality enhancement, and lifecycle management of polysaccharide conjugate vaccines.

    2025 08 v.38 [Abstract][OnlineView][Download 898K]

  • Considerations on nonclinical pharmacodynamics studies of prophylactic mpox vaccine

    YIN Huajing;WU Shuang;WANG Yin;YIN Maoshan;LI Zheng;YU Bing;SUN Tao;Center for Drug Evaluation, National Medical Products Administration;

    Mpox, previously known as monkeypox, is a zoonotic disease caused by the mpox virus(MPXV), which has shown a trend of spreading globally in recent years and poses a threat to public health security. To address the potential mpox epidemic, the development of vaccine has become crucial. Nonclinical pharmacodynamics studies are an important part of vaccine development, which evaluate the immunogenicity and protective effect of vaccines in animal models, providing key evidence of efficacy to support clinical trials. This article will explore the key technical points of nonclinical pharmacodynamics studies for mpox vaccines, in combination with the nonclinical efficacy studies of similar products and the technical documents recently released, in order to provide some reference for the development of such products.

    2025 08 v.38 [Abstract][OnlineView][Download 856K]
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@2012 Editorial Department of "Chinese Journal of Biologicals"