2025年05期目次

  • Adsorption properties of amorphous aluminum hydroxyphosphate sulfate adjuvant and aluminum hydroxide adjuvant for recombinant hepatitis B surface antigen

    JIANG Ying;HAO Yueru;REN Huiwen;JI Wenheng;ZHAO Yuxiu;WANG Yunyang;Beijing Institute of Biological Products Co., Ltd.;

    Objective To investigate the adsorption mechanism of amorphous aluminum hydroxyphosphate sulfate(AAHS)adjuvant and aluminum hydroxide adjuvant on recombinant hepatitis B surface antigen(HBsAg) and the effect of differentadsorption rates on the titer of hepatitis B.MethodsThe Langmuir adsorption equation was used to calculate the maximumadsorption capacity of HBsAg on the surface of AAHS and aluminum hydroxide adjuvants at different concentrations ofsodium chloride(NaCl) and ethylene glycol. Combined with the influence of different concentrations of phosphate on theadsorption effect of HBsAg, the main forces affecting the adsorption of HBsAg and two aluminum adjuvants were analyzed,and the difference between the two adjuvants was also compared. ED_(50)was used to evaluate the immune effect of sampleswith different phosphate concentrations, and the relationship between different adsorption rates and titers was analyzed.ResultsDifferent concentrations of NaCl and ethylene glycol did not affect the adsorption rate of HBsAg, and the adsorptionrate decreased with the increase of phosphate concentration. The titer of samples with lower adsorption rate was still withinthe qualified standard range.ConclusionHBsAg is mainly adsorbed on the surface of AAHS and aluminum hydroxide adju-vants through ligand exchange, and hepatitis B titer may not require high antigen adsorption rate in vitro.

    2025 05 v.38 [Abstract][OnlineView][Download 940K]

  • Evaluation of humoral and cellular immunity of mice immunized with inactivated SARS-CoV-2 vaccine against Delta strain

    XU Kangwei;XU Li ;LU Xu;LI Jing ;GE Xiaoqin ;WANG Kaiqin;LI Changgui;QUAN Yaru;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 evaluate the humoral and cellular immune effects of BALB/c mice immunized with prototype inac-tivated SARS-CoV-2 vaccine against Delta strain virus, and to provide a reference for evaluating the protective effect ofexisting vaccines against variant strains and developing safer and more effective vaccines.MethodsFemale BALB/c micewere intraperitoneally immunized with inactivated SARS-CoV-2 vaccine twice at an interval of 14 days, and mice immu-nized with PBS were used as control group, with 10 mice in each group. Serum samples were collected on the 7th, 14th,21st, 28th, 35th and 42nd days after the initial immunization. The binding antibody titers against the S and N proteins ofDelta strain virus were detected by indirect ELISA, and the neutralizing antibody titers against Delta strain virus weredetected by microneutralization test. On the 42nd day after the initial immunization, the spleen of mice was taken for Elispottest to evaluate the cellular immunity level.ResultsThe S protein-binding antibody was detected 7 days after the initialimmunization, and the antibody titer increased further after the boosting immunization, reaching a geometric mean titer(GMT) of 89 144 on the 21st day. However, the level of N protein-binding antibody was lower after the initial immunization,and increased rapidly after the boosting immunization, which was comparable to the level of S protein-binding antibody. Onthe 7th and 14th day after the initial immunization, the seroconversion numbers of neutralizing antibodies were 4/10 and 8/10, while all the antibodies showed seroconversion after the boosting immunization, with the GMT of neutralizing antibodiesof 391. On the 42nd day after the initial immunization, the average spots of IFNγ and IL-2 in the vaccine group were signifi-cantly more than those in the control group(t = 8. 094 and 13. 08, respectively, each P < 0. 000 1).ConclusionTwo timesimmunization with inactivated SARS-CoV-2 vaccine can effectively stimulate humoral and cellular immunity against Deltastrain virus in mice.

    2025 05 v.38 [Abstract][OnlineView][Download 862K]

  • Stability of tick-borne encephalitis inactivated vaccine(0. 5 mL/dose)with pre-potting syringe

    CHEN Nana;MIAO Hui;GONG Xiaotang;ZHANG Dongxue;LIU Zhencheng;SUN Hongliang;ZHANG Ying;LI Jingliang;Changchun Institute of Biological Products Co., Ltd.;

    Objective To evaluate the stability of tick-borne encephalitis inactivated vaccine(0. 5 mL/dose) with pre-pottingsyringe in order to ensure the effectiveness and safety of the product. Methods The optimized stock solution of tick-borneencephalitis inactivated vaccine(0. 5 mL/dose, pre-potting syringe) was stored at(5 ± 3) ℃ for 120 days. The indicators ofsterility, antigen content, protein content, bovine serum albumin residue and hamster kidney cell protein residue weredetected and compared with those of vaccine before optimization(0. 1 mL/dose, vial). The long-term stability(0-33 monthsat 2-8 ℃), accelerated stability [0, 3, 6 months at(25 ± 2) ℃] and accelerated destruction stability [0, 7, 14 days at(37 ±1) ℃] of the optimized tick-borne encephalitis inactivated vaccine were investigated according to the relevant requirements ofTechnical Guidelines for Stability Research of Biological Products, including appearance, titer, aluminum content, osmoticmolar concentration and other indicators. Results Before and after optimization, all indicators of the stock solution of tick-borne encephalitis inactivated vaccine were qualified. There was no statistically significant difference in bovine serumalbumin residue and hamster kidney cell protein residue(F = 3. 33 and 0. 61, P = 0. 077 and 0. 627, respectively), but theF P differenceofproteincontentwasstatisticallysignificant(=38.07,<0.05),whileallofthemwerewithinthestandard rangeof ≤ 80 μg/mL. After optimization, the indicators of long-term stability, accelerated stability and accelerated destructionstability of the finished product of tick-borne encephalitis inactivated vaccine were all qualified. In the tests of long-termstability, respectively the vaccine titer after optimization was significantly higher than that before optimization(F = 62. 32,P < 0. 05); the differences of osmotic pressure molar concentration were statistically significant(F = 40. 47 and 12. 50 respec-tively, each P < 0. 05), but the fluctuation was controlled within the standard range of 240-320 mOsmol/kg. Conclusion Tick-borne encephalitis inactivated vaccine(0. 5 mL/dose) with pre-potting syringe has good stability under the conditions of long-term storage, transportation and short-term overtemperature, and its quality meets the standard.

    2025 05 v.38 [Abstract][OnlineView][Download 846K]

  • Cross-neutralization effect of multivalent recombinant receptor-binding domain dimer protein vaccine on SARS-CoV-2 variant strains

    CAO Qing;MEN Yunzheng;ZHANG Yanling;LIU Xiaoya;RONG Huan;SHE Guangbiao;WU Changwei;HUANG Enqi;Anhui Zhifei Longcom Biopharmaceutical Co., Ltd.;

    Objective To evaluate the cross-neutralization effect of multivalent recombinant receptor-binding domain(RBD)dimer protein vaccine against SARS-CoV-2 variant strains, so as to provide an experimental basis for the development ofbroad-spectrum SARS-CoV-2 vaccines. Methods Twenty-two female SD rats were randomly divided into ZF2202(B) group(8 rats immunized with quadrivalent recombinant RBD dimer protein vaccine), ZF2202(A) group(6 rats immunized withbivalent recombinant RBD dimer protein vaccine) and normal saline control group(8 rats). The rats were immunized intra-muscularlyat0,21and42days,respectively,withadoseof0.25mL/rat.Fourteendaysafterthelastimmunization,theneutralizingantibodies against Delta, BA.4/5, BQ.1.1, XBB.1, EG.5 and BA.2.86 in serum of rats in ZF2202(B) and ZF2202(A) groupswere detected by pseudovirus neutralization assay, and the secretion of IFNγ and IL-4 in spleen lymphocytes of rats in ZF2202(B)and normal saline control groups were analyzed by ELISpot. Results The levels of neutralizing antibodies againstpseudovi-ruses BA.4/5, BQ.1.1, XBB.1, EG.5 and BA.2.86 in serum of rats in ZF2202(B) group were significantly higher than those inZF2202(A) group(U = 3, 1, 1, 1, 0, P = 0. 004 7, 0. 001 3, 0. 001 3, 0. 001 3, and 0. 000 7, respectively). Compared withnormal saline group, the number of specific spots of IFNγ secreted by spleen lymphocytes in ZF2202(B) group increased signifi-cantly(U = 4, P = 0. 001 4), and the number of IL-4 specific spots also increased significantly(t = 3. 775, P = 0. 002 1).Conclusion Both ZF2202(A) and ZF2202(B) provide cross-protective immunity against SARS-CoV-2 variants, whileZF2202(B)exhibitsbetterprotectiveeffectthanZF2202(A)ontheepidemicstrains,andexhibitsabroaderspectrumofefficacy.

    2025 05 v.38 [Abstract][OnlineView][Download 863K]

  • Culture conditions optimization and biological characteristics analysis of MU-K3 human embryonic kidney cells

    JIN Liwu;ZHANG Zhenyu;YANG Yawen;TIAN Ling;TANG Xiaolin ;WANG Jiamin;QIAO Zilin;Gansu Tech Innovation Center of Animal Cell, Biomedical Research Center, Northwest Minzu University;

    Objective To optimize the culture conditions and investigate the biological characteristics of MU-K3 humanembryonic kidney cells, so as to lay a foundation for the study of related biological functions of human embryonic kidneycells. Methods MU-K3 cells were cultured with different media(DMEM, MEM, M199, DME/F12), serum types and concen-trations [10%, 8%, 6%, 4% newborn bovine serum(NBS) and 10% fetal bovine serum(FBS)], and passage ratios(1∶2, 1∶4,1∶8). The optimum culture conditions were determined with the maximum proliferation concentration, doubling time andspecific growth rate as evaluation indexes. The cell cycle and apoptosis of MU-K3 cells were detected by flow cytometry. MU-K3cells were subcultured continuously under the best conditions until the cells could not form a dense monolayer and died ofsenescence. The limited culture generations were determined, and the senescence of cells was analyzed by β-galactosidasestaining. The expression of vimentin in cells was measured by immunofluorescence assay. The chromosome karyotype wasanalyzed by Giemsa staining to evaluate genetic stability. Results The optimum culture conditions for MU-K3 cells were asfollows: cultured in DMEM medium containing 10% FBS and subcultured in a ratio of 1∶4. In P9 MU-K3 cells, the proportionof S phase cells was 23. 34%, and the apoptosis rate was 10. 61%. With the increase of passages(P17), the proliferationability decreased(the proportion of S phase cells was 12. 22%, and the apoptosis rate was 6. 38%). The limited culture genera-tion of MU-K3 cells was P19, the activity of β-galactosidase in the cells increased gradually with the increase of generations,and the cells showed senile phenotype. The MU-K3 cells were fibroblast-like, with the number of chromosomes of 2 n = 46,belonging to normal human diploid karyotype. Conclusion The culture conditions of MU-K3 cells were optimized. MU-K3cells are normal human diploid cells, with stable proliferation ability and genetic characteristics under the optimum cultureconditions, which can be used as an ideal cell model for kidney development and disease research.

    2025 05 v.38 [Abstract][OnlineView][Download 1246K]

  • Expression and enzymatic properties of urate oxidase from Candida utilis in Lactococcus lactis

    SHI Ping;LIU Yuansen;PENG Jingjing;LIAO Jiajun;WU Qingling;GUO Dejun ;WANG Chenghua;College of Light Industry and Food Engineering, Guangxi University;

    Objective To express urate oxidase(UOX) from Candida utilis in lactic acid bacteria and study its enzymaticproperties, so as to provide technical supports for the application and development of UOX.MethodsThe optimized UOXgene was synthesized and cloned into the vector pTRKH2 to construct the recombinant expression plasmid, which was thenelectroporated to competent Lactococcus lactis NZ9000 to screen the recombinant strain and express the recombinant UOX(rUOX) with His-tag. Using UOX enzyme activity as an index, the fermentation conditions of recombinant bacteria were opti-mized. The rUOX was purified by nickel ion metal chelate chromatography, and its enzymatic properties were characterized.ResultsThe optimum expression conditions of the recombinant strain were as follows: fermentation temperature 37 ℃,fermentation time 12 h, and initial pH 6 of fermentation medium, respectively, and the rUOX enzyme activity reached(44. 57 ±0. 43) U/mL. The relative molecular mass of purified rUOX was about 35 000, consistent with the theoretical relative molecu-lar mass, and the specific activity was 17. 48 U/mg. The optimum pH and temperature of rUOX were 11. 5 and 45 ℃, andunder the optimal conditions, the K_m, V_(max)and k_(cat)values of rUOX were 5. 72 mmol/L, 71. 25 μmol/(L·min) and 95. 76/s,respectively, with uric acid as the substrate. Various metal ions and chemical reagents had inhibitory activity on rUOX, whileTween 20 exhibited obvious activation activity on rUOX.ConclusionThe recombinant expression of UOX in lactic acidbacteria is feasible, and its enzyme activity is relatively stable, which provides an experimental basis for the development andapplication of UOX under alkali-tolerant conditions.

    2025 05 v.38 [Abstract][OnlineView][Download 1010K]

  • Preparation of monoclonal antibodies against human leukocyte immunoglobulin like receptor B2 and preliminary identification of their activity in vitro

    FENG Yanping;HU Yuqing;WU Guojin ;LIANG Jiabei;NING Jinying ;LI Li;College of Pharmacy, Xinjiang Medical University;

    Objective To prepare monoclonal antibodies against human leukocyte immunoglobulin like receptor subfamilyB2(LILRB2) by hybridoma technique and preliminarily identify their activity in vitro, so as to provide a new strategy for targeting“cold”tumor therapy.MethodsThe extracellular region of human LILRB2 protein was expressed, purified by affinity chro-matography, and then used to immunize BALB/c mice to prepare anti-LILRB2 monoclonal antibodies with high specificityand affinity. The variable regions of light chain and heavy chain of mouse monoclonal antibodies were inserted into an expres-sion vector containing human constant regions by DNA recombination technique to produce chimeric antibodies, and theactivity in vitro was identified by ELISA, flow cytometry and bio-layer interferometry(BLI).ResultsSeven anti-LILRB2monoclonal antibodies with high purity were successfully prepared by hybridoma technique, all of which could recognizeLILRB2 protein with high specificity. Among them, 11C6-8, 43H7-2 and 53A6-11 had superior performance, with the EC_(50)values of 0. 272 3, 0. 431 8 and 0. 344 0 μg/mL, respectively. The chimeric antibodies obtained by humanized modificationexhibited higher binding ability to LILRB2 and effectively blocked the binding of LILRB2 to its ligand, human leukocyteantigen-G(HLA-G), among which the IC_(50)values of 11C6-8 and 53A6-11 were 0. 429 2 and 0. 283 6 μg/mL.ConclusionThe specific monoclonal antibodies against LILRB2 were successfully prepared, and expected to be developed into new anti-tumor immune antibody drugs, which provides a new idea for the development of therapy strategy targeting tumors.

    2025 05 v.38 [Abstract][OnlineView][Download 1206K]

  • Antitumor activity of filtrate of a novel Staphylococcus aureus with incomplete hemolytic phenotype

    LI Xinyu;LIU Ying ;LI Xin;YAO Jie;ZHOU Qiang;TANG Wei;Department of Clinical Laboratory, the Second Affiliated Hospital of Anhui Medical University;

    Objective To investigate the antitumor activity of the filtrate of a novel Staphylococcus aureus with incompletehemolytic phenotype(SIHP), and to provide references for screening new antitumor drugs.MethodsThe epidemiologicalcharacteristics of the novel SIHP were summarized and analyzed. The hemolytic phenotypes of the novel SIHP, classic Staphy-lococcus aureus with complete hemolytic phenotype(SCHP) and quality control strains ATCC29213 and ATCC25923 weredetected on blood agar plate(BAP) by three-point method. Staphylococcus aureus filtrate was prepared by inoculating LBbroth, centrifuging and filtering after fermentation. The Staphylococcus aureus filtrate and its diluents with RPMI1640 ofdifferent volume fractions(1∶1, 1∶3, 1∶7) were co-incubated with human non-small cell lung cancer HCC827 cells, themorphological changes were observed, and the cells were classified and counted under a microscope.ResultsThe clinicalspecimen isolation rate of the novel SIHP was only 2. 01%, and its hemolytic phenotype on BAP was unique with strongrecognition. Compared with the classical SCHP and quality control strain ATCC25923, the novel SIHP filtrate showed signifi-cantly stronger tumor cytotoxicity, and damaged HCC827 cells in a concentration-dependent manner, leading to the release of cell contents, balloon-like degeneration of cell membrane or bare nucleus-like degeneration of cells.ConclusionThe novel SIHP may be a new subspecies of Staphylococcus aureus with a unique genetic background, and the filtrate contains highly effective antitumor active substances, which can provide a material basis for the screening of new antitumor drugs.

    2025 05 v.38 [Abstract][OnlineView][Download 1001K]

  • Inhibitory effect of norcantharidin on non-small cell lung cancer by targeting BCL-2/P53 proteins

    DONG Yu;FAN Kaiqi;Pharmacy Department of Jiamusi Central Hospital;

    Objective To analyze the expression of BCL-2 and P53 proteins in non-small cell lung cancer(NSCLC) and theinhibitory effect of norcantharidin(NCTD) on NSCLC by targeting the expression of these two key proteins, so as to provideexperimental evidence for developing new therapeutic strategies for NSCLC.MethodsBioinformatics methods were used toanalyze the expression of BCL-2 and P53 in cancer, especially lung cancer, and their correlation with the survival rates ofpatients. The effects of NCTD on proliferation, apoptosis and the expression of BCL-2 and P53 proteins in NSCLC cells weremeasured by CCK-8, flow cytometry and Western blot, respectively.ResultsBCL-2 and P53 proteins were found to behighly expressed in various tumors, with a more pronounced expression in lung cancer(t = 3. 810, P < 0. 05). The expressionof BCL-2 protein was significantly correlated with the survival rates of lung cancer patients(t = 3. 235, P < 0. 05). Theaffinity of NCTD to BCL-2 protein was significantly higher than that to P53 protein. NCTD inhibited the proliferation ofNSCLC cells, promoted the apoptosis, and significantly reduced the expression of BCL-2 and P53 proteins(t = 3. 25 and3. 45, respectively, each P < 0. 05).ConclusionBCL-2 and P53 play a crucial role in the development of NSCLC, andNCTD can effectively inhibit the expression of these two proteins, thereby inhibiting the proliferation of NSCLC cells. Thisstudy provides an experimental basis for developing new therapeutic strategies for NSCLC, and also lays a foundation forfurther research on NCTD as a potential anti-tumor drug.

    2025 05 v.38 [Abstract][OnlineView][Download 1080K]

  • Safety analysis of different types of hepatitis B vaccines in Fujian Province, 2019-2023

    LIN Zhiqiang;XIAO Jianxiong;CHEN Zhifei;WU Ruihong;PAN Weiyi;WANG Qin;Fujian Provincial Center for Disease Control and Prevention;

    Objective To analyze the epidemiological characteristics of adverse events following immunization(AEFI) ofthree types of hepatitis B vaccines(CHO, Saccharomyces cerevisiae and Hansenula polymorpha) in Fujian Province from 2019to 2023, and evaluate their safety. Methods The AEFI case data and vaccination data of hepatitis B vaccines reported inFujian Province from 2019 to 2023 were collected through the Chinese Immunization Planning Information ManagementSystem. The occurrence characteristics, incidence rates and other indicators of AEFI related to hepatitis B vaccines wereanalyzed by using descriptive analysis methods. Results In Fujian Province from 2019 to 2023, there were 677 reportedcases of AEFI related to hepatitis B vaccines, with an AEFI reported incidence of 7. 44 per 100 000 doses. The male-to-female ratio was 1. 20∶1, and the age distribution was mainly in the group of less than 1 year old. The reported incidence ofcommon adverse reactions was 6. 24 per 100 000 doses, and the reported incidence of rare adverse reactions was 0. 96 per100 000 doses. Rare vaccine reaction cases were mainly anaphylactic reaction. There were statistically significant differencesin the reported incidence of common adverse reactions and allergic skin rashes among the three types of hepatitis B vaccines(χ~2= 111. 587 and 13. 284, respectively, each P < 0. 05). Conclusion The reported incidence of AEFI related to hepatitisB vaccines in Fujian Province has been rare from 2019 to 2023, falling within the expected range of WHO, indicating goodsafety of the vaccines.

    2025 05 v.38 [Abstract][OnlineView][Download 849K]

  • Development and verification of a crystal violet-methylcellulose plaque reduction neutralization test for detection of neutralizing antibody titer of tick-borne encephalitis virus

    ZHAO Dongmei;QIU Lu;ZHANG Xu;GAO Xianxian;ZHOU Yangyang;YU Lili;LI Shuai;Biosafety Level-3 Laboratory, Changchun Institute of Biological Products, Co., Ltd.;

    Objective To develop a crystal violet-methylcellulose plaque reduction neutralization test for detecting the titerof neutralizing antibody against tick-borne encephalitis virus(TBEV), and to verify and preliminarily apply the method, so asto provide a reliable method for TBEV antibody products in the detection of live virus neutralizing titer.MethodsOrthogo-nal experimental design was used to determine the important parameters involved in the experiment, namely, the best experi-mental conditions of virus culture time, virus-cell incubation time, neutralization temperature and methylcellulose concentra-tion. According to the experimental parameters determined above, the method was verified for specificity, relative accuracy,linearity, repeatability and intermediate precision. The titer of neutralizing antibody in serum of population immunized withinactivated tick-borne encephalitis vaccine was detected by the developed method.ResultsThe optimum conditions were asfollows: virus culture time of 5 d, virus-cell incubation time of 1. 5 h, neutralization temperature of 37 ℃ and methylcelluloseconcentration of 3%. Under these conditions, clear round plaques were formed, which was convenient for counting. Neutrali-zing antibodies were detected in both diluted test group and accelerated destruction test group in specificity verification withthe detection results not less than 10, while the results of impurity group were less than 10. In the relative accuracy verifica-tion, the relative bias of each titer was 0-10%, all in the range of ± 12%, and the linear regression correlation coefficient(r)was 0. 999 3, which was greater than 0. 98, In the verification of repeatability and intermediate precision, the geometric coef-ficients of variation(GCVs) of each group of samples were between 1. 5% and 4. 1%, not more than 20%. The serum neutrali-zing antibody titers of the population immunized with inactivated tick-borne encephalitis vaccine were less than 10 beforeinitial immunization, and not less than 10 after initial immunization and before and after booster immunization, and theserum neutralizing antibody increased after booster immunization.ConclusionThe developed TBEV neutralizing antibodytiter detection method has good durability, specificity, relative accuracy, linearity and precision, and the results are easy todetermine, which can be used for TBEV neutralizing antibody titer detection.

    2025 05 v.38 [Abstract][OnlineView][Download 923K]

  • Establishment of atomic absorption spectrophotometry for determination of calcium content in plasma-derived FⅧ products and evaluation of uncertainty

    ZHANG Ting;ZHOU Changming;SHEN Cong;SHAO Tianshu;DING Rui;Beijing Institute for Drug Control (Beijing Center for Vaccine Control), NMPA Key Laboratory for Research and Evaluation of Generic Drug, Beijing Key Laboratory of Analysis and Evaluation on Chinese Medicine;

    Objective To establish an atomic absorption spectrophotometry for the determination of calcium content in humanplasma-derived FⅧ(pdFⅧ) products, and to evaluate the uncertainty of the method, so as to provide a reliable method forthe determination of calcium content in blood products. Methods After adding lanthanum chloride solution as ionizationinhibitor to pdFⅧ, the calcium content was detected by atomic absorption spectrophotometry. The gas flow rate(1. 2, 1. 4,1.6,1.8,2.0L/min),burnerheight(9,11,12,13mm),lampcurrent(100%,90%,85%,80%),andslitwidth(0.2,0.5,1.0nm)were optimized, and the method was verified for specificity, linearity range, limit of detection(LOD), limit of quantitation(LOQ),accuracy,precisionandstability.ThecalciumcontentinsixbatchesofpdFⅧsampleswasdeterminedbytheestablishedmethod.Theuncertaintyofthemethodwasevaluated,andtakingpdFⅧsampleofbatchnumber202404026asanexample,thecalcium content was determined, and the effects of various sources of uncertainty on the detection results were determined.Results The optimum gas flow rate was 1. 8 L/min, the combustion gas height was 12 mm, the lamp current was 85%,and the slit width was 0. 5 nm. Lanthanum chloride did not interfere with the determination of calcium content in pdFⅧ. Inthe concentration range of 1. 0-5. 0 μg/mL, the calcium working standard solution had a good linear relationship with A_(422.7),with the regression equation of: y = 0. 016 6 x-0. 000 8, r = 0. 998 7. The LOD was 0. 127 2 μg/mL, and the LOQ was0. 385 5 μg/mL. The recovery rates of low, medium and high concentration spiked samples were all in the range of 85%-110%. The RSDs of precision and stability verification were both less than 2%. The calcium content of six batches of pdFⅧsamples was not more than 50 μg/mL. The uncertainty was mainly derived from the preparation of standard solution. Thecalcium content of pdFⅧ sample with batch number 202404026 was(42. 11 ± 1. 736 1) μg/mL, which met the requirement(≤ 50 μg/mL), indicating that the sources of uncertainty showed no influence on the determination results of calcium concen-tration. Conclusion The established atomic absorption spectrophotometry has good accuracy, precision and stability, andthe reliability of detection results is ensured by uncertainty evaluation, which can be used for the accurate determination ofcalcium content in pdFⅧ.

    2025 05 v.38 [Abstract][OnlineView][Download 892K]

  • Analysis of host cell protein residues in bevacizumab by Nano-UPLC-MS/MS and ELISA

    LU Jingjing;HUO Linan;Medicine&Pharmacy Research Center, Binzhou Medical University;

    Objective To analyze the host cell protein(HCP) residues in bevacizumab injection and its downstream productsby nano ultra-performance liquid chromatography-tandem mass spectrometry(Nano-UPLC-MS/MS) and enzyme-linked immu-nosorbent assay(ELISA).MethodsThe samples were directly treated with trypsin, and the undigested antibodies wereprecipitated by heat treatment, which were then detected by Nano-UPLC connected with Q-Exactive Mass Spectrometer. Thecollected peptide information was compared with the total protein library of Chinese hamsters by MaxQuant software, and theHCP types were identified. The absorbance values of samples were detected by double-antibody sandwich ELISA, and theHCP content was calculated according to the four-parameter curve.ResultsAfter purification by downstream process, thelevels and types of HCPs decreased significantly. The content of HCPs in injection was 7 ppm, including 20 types, amongwhich 5 types of persistent residual HCPs were detected, and 4 types of high-risk HCPs were not detected. Compared withthe original drug, the HCP residues of the two drugs were both less than or equal to 15 ppm, and there were differences inHCP types due to different purification processes.ConclusionThe combination of Nano-UPLC-MS/MS and ELISA providesa comprehensive analytical strategy for the characterization and quantification of HCP, especially the types and distributionof persistent residual HCP and high-risk HCP, which can provide a reference for the process optimization, comparabilityresearch and safety risk assessment of products.

    2025 05 v.38 [Abstract][OnlineView][Download 911K]

  • Establishment and verification of dual TaqMan probe qPCR detection method for target gene copy number of genetically attenuated pertussis vaccine strains

    BU Xiangli;SUN Yeting ;ZHANG Lizhi ;ZHAO Ran ;LIN Jisheng ;LI Jing ;WANG Miao;Shenyang Pharmaceutical University;

    Objective To establish a dual TaqMan probe-based qPCR method for determining the copy number of the geneti-cally modified pertussis toxin(PT) gene in detoxified pertussis vaccine strains, and to identify optimal reference genes forevaluating the genetic stability of modified strains during serial passage.MethodsSpecific primers and probes of modifiedgene PT and multiple housekeeping genes were designed. The optimal reference genes were screened by GeNorm, Norm-Finder, BestKeeper and Delta CT based on dual TaqMan probe qPCR. The copy number of PT gene of genetically detoxifiedpertussis vaccine strain was calculated by relative quantitative analysis. The specificity, linearity and repeatability of theestablished method were verified. The genomic DNA of P1, P3, P5, P10 and P15 strains used in the production of geneticallydetoxified pertussis vaccine was used as templates, and the passage stability of the strains was evaluated by using the estab-lished dual TaqMan probe qPCR method.ResultsThe stability of candidate reference genes was analyzed using GeNorm,NormFinder, BestKeeper, and Delta CT software, with Rpob identified as the most stable reference gene. Both the target PTgene and the reference Rpob gene were amplified in standard strain 58003. A good linear correlation(R~2≥ 0. 990) wasobserved between Ct values and template quantities within the concentration range of 0. 001 to 10 ng/μL. For P1 generationstrains, the 2~(-△△Ct)values from six replicate assays consistently ranged between 0. 87 and 1. 25, with an RSD of less than 20%.All tested generations exhibited single-copy PT target genes, demonstrating good genetic stability during serial passages.ConclusionThe established dual TaqMan probe qPCR assay demonstrates good primer specificity and reproducibility,making it suitable for evaluating the genetic stability of bacterial strains during serial passages in the production of geneti-cally detoxified pertussis vaccines, thereby further evaluating the quality of strains.

    2025 05 v.38 [Abstract][OnlineView][Download 1073K]

  • Research progress on mechanism of mesenchymal stem cells in treatment of deep vein thrombosis

    ZHANG Bo;SU Yuan;HA Xiaoqin;School of Basic Medicine, Gansu University of Traditional Chinese Medicine;

    Thrombotic disease represents a significant peril to human life and well-being, with the highest clinical incidenceacross all maladies, rendering it a paramount focus of contemporary medical inquiry. Deep vein thrombosis(DVT), as asubtype of thrombotic diseases, has exhibited an escalating prevalence in recent years. Presently, there exists a paucity ofclinical interventions for DVT, predominantly palliative in nature rather than curative. Hence, the quest for novel methodolo-gies to effectively prevent and treat DVT has surgent practical significance. Studies have demonstrated the pivotal role ofmesenchymal stem cells(MSCs) in the management of DVT, highlighting their significant potential for extensive clinicalapplications. This paper endeavors to comprehensively expound up on the action mechanism of MSCs in DVT treatment withthe aim of furnishing novel insights and methods for clinical management.

    2025 05 v.38 [Abstract][OnlineView][Download 871K]

  • Epidemiological status, related diseases and vaccine advances of non-typeable Haemophilus influenzae

    HU Nan;WANG Cai;CHANG Yueli;ZHANG Yutuo;Institute of Pathogenic Biology and Immunology, Hebei North University;

    Non-typeable Haemophilus influenzae(NTHi) is a non-encapsulated, Gram-negative coccobacillus that commonlycolonizes in human upper respiratory tract and can cause invasive infections in immunocompromised hosts. In recent years,with the widespread implementation of Haemophilus influenzae type b(Hib) vaccination, NTHi has emerged as the predominantpathogenic strain of Haemophilus influenzae infections globally. Notably, the clinical isolation rate of NTHi has shown aprogressive increase worldwide, accompanied by rising resistance to β-lactams and other antibiotics. NTHi frequently formspolymicrobial infections with pathogens such as Streptococcus pneumoniae and Moraxella catarrhalis, and contributes tovarious diseases such as otitis media, meningitis, and acute exacerbations of chronic obstructive pulmonary disease(COPD)through virulence mechanisms including biofilm formation and immune evasion. At present, antibiotics remain the maindrugs for NTHi infection management, while increasing antimicrobial resistance has driven vaccine development as a criticalresearch priority. Promising vaccine candidates based on antigens such as outer membrane proteins and adhesins, havedemonstrated favorable immunogenicity and protective efficacy, but further optimization is required to enhance cross-protectiveimmunity. In this paper, the epidemic status, related diseases and vaccine research progress of NTHi are summarized, aimingto provide references for the prevention and control of NTHi infection.

    2025 05 v.38 [Abstract][OnlineView][Download 906K]

  • Research progress of type Ⅲ interferons

    GAO Jian;ZHOU Baisong;LIU Yulin;LIU Jinghui;The second Research Laboratory, Changchun Institute of Biological Products Co., Ltd.;

    Interferons(IFNs) are a family of glycoproteins with antiviral, antitumor, and immunomodulatory activities, classi-fied into type Ⅰ, Ⅱ, and Ⅲ based on structural and receptor specificity. Among these, type Ⅲ IFNs(IFNλs), including IFNλ1(IL-29),IFNλ2(IL-28a),andIFNλ3(IL-28b),exhibittissue-specificreceptordistribution,predominantlyexpressedinepithelialcells and select immune cells, thereby demonstrating reduced systemic side effects compared to typeⅠIFNs. Mechanistically,IFNλs exert their antiviral effects through JAK/STAT signaling pathway inducing the expression of interferon-stimulatedgenes(ISGs), and inhibit tumor cell proliferation, promote apoptosis and regulate immune responses. Recent advances inrecombinant IFNλs therapeutic development include polyethylene glycol(PEG) modified IFNλ1, a longacting formulationcurrently in phaseⅢclinical trials for hepatitis C, hepatitis D, and COVID-19. In this paper, the structures and receptors,signaling pathway and regulation, biological functions and research progress of IFNλs are summarized in order to provideinsights for the development of IFNλs related products.

    2025 05 v.38 [Abstract][OnlineView][Download 1019K]

  • Research progress of new adjuvants in development of SARS-CoV-2 vaccines

    SU Linjia;XU Fangjingwei;ZHANG Yuntao;Research and Development Management Department, China National Biotec Group Company Limited;

    Vaccination is the most cost-effective strategy to prevent and control SARS-CoV-2 infection, which plays an importantrole in maintaining global public health and ensuring human health. Adjuvants, as crucial components in vaccine development,can make up for the potential antigen immunogenicity deficiencies by enhancing the speed, intensity and persistence ofimmune response, and significantly improve the efficacy of vaccines, being a pivotal role in improving the immune effect ofSARS-CoV-2 vaccines. There are many types of adjuvants, which are suitable for different types of vaccines. For example,the inactivated SARS-CoV-2 vaccine mostly uses aluminum adjuvant, the recombinant protein vaccines adapt to compositeadjuvants, and the mRNA vaccines rely on lipid nanoparticle(LNP) delivery platform. In this paper, the classification andmechanism of adjuvants, their application in vaccines and new ideas for development of broad-spectrum SARS-CoV-2 vaccineadjuvants were reviewed, providing references for preparing innovative adjuvants with good safety and effectiveness, so as tocontinuously improve the immune efficacy of SARS-CoV-2 vaccines and cope with the continuous variation of SARS-CoV-2.

    2025 05 v.38 [Abstract][OnlineView][Download 850K]

  • Evolution, current situation and reflections of biological reference materials' technical guidelines and management regulations in WHO and in China

    WANG Yiping;MAO Qunying ;WANG Xiaojuan ;LIANG Zhenglun;The Center for Reference Material and Standardization, National Institutes for Food and Drug Control;

    Reference material is a kind of criteria“ruler”for the quality control and evaluation of biological products. WorldHealth Organization(WHO) technical reports and guidelines, China's management regulations about biological referencematerials and Chinese Pharmacopoeia play an important guiding role in the development,distribution and use of standards. Inorder to meet the needs of the development of the biological product industry, WHO guidelines and Chinese national manage-ment regulations about biological reference materials have been revised for several times. This paper summarizes the evolu-tion and current situation of the definition, classification and technical requirements for WHO international biological stan-dards and national biological standards in China, aiming to further improve the preparation and application of national biologi-cal reference materials in China.

    2025 05 v.38 [Abstract][OnlineView][Download 844K]

  • Key points and defect analysis in bioproduct quality control:Insights from research and production site inspections

    YANG Jingpeng;SHEN Hong ;CAO Yi;Center for Food and Drug Inspection of NMPA;

    As biopharmaceutical technology continues to advance, the global market for bioproducts is growing, and the inno-vation and development of these products have become a crucial component of the Healthy China strategy. In this setting,ensuring the quality of bioproducts is especially critical. Quality control is vital not only for public health and safety but alsofor the industry′s sustainable growth and international competitiveness. Quality control laboratories, as the cornerstone of acompany′s quality system, play a key role in ensuring that products adhere to high standards through meticulous testing andanalysis. This process helps protect public health, drive industry advancement, and boost international competitiveness. Thisstudy seeks to systematically examine the verification elements and common deficiencies in bioproduct quality control labora-tories, identifying key control points and frequent issues within the bioproduct quality management system. It offers scientificsupport for regulatory bodies and aids companies in building and maintaining high-quality management systems to ensure thesafety and efficacy of bioproducts, safeguard public health, foster ongoing industry improvement and technological innova-tion, and raise the overall industry standard.

    2025 05 v.38 [Abstract][OnlineView][Download 814K]
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@2012 Editorial Department of "Chinese Journal of Biologicals"