THE SPECIFIC PHOTODYNAMIC EFFECTS OF MONOCLONAL ANTIBODIES CONJUGATED WITH HEMATOPORPHYRIN DERIVATIVE ON GASTRIC CANCER IN VITRO AND IN VIVO

Murine monoclonal antibody (MoAb) BB4.3, raised against the human gastric cancer cell line BGC823, was puriffied with Protein A-Sepharose CL-4B affinity chromatography and identified as IgG2a. It was then conjugated with a hematoporphyrin derivative (HPD) by using carbodiimide. The qualitative analysis of this conjugate showed that the amount of free HPD was negligible and there were no IgG aggregates among the conjugates. The conjugate retained both the antibody and photochemical activity of HPD.In vitro, the phototoxic effect of this HPD-BB4.3 conjugate on target cells was about 15 times higher than that of free HPD. The quality of selective photocytotoxicity was proven by the greater cytotoxi-city this conjugate showed than that of corresponding normal mouse IgG (NIgG) conjugated with HPD. It showed less cytotoxicity to colon cancer cell line B-80 (negative reaction to MoAb BB4.3) than to BGC825. Moreover, its cytotoxicity to BGC823 cells could be blocked specifically by excess BB4.3 antibody, but not by another MoAb 3G9, which combines with BGC823 at different binding sites from MoAb BB4.3.Nude mice inoculated with 2 × 10- BGC823 cells were given HPD-BB4.3, HPD, HPD-NIgG, HPD plus BB4.3 and PBS, respectively then exposed to light. Four out of six animals treated with the HPD-BB4.3 conjugate remained tumor-free for a long period. Although two developed tumors, there was a significant difference between the HPD-BB4.3-treated group and all the control groups in tumor induction time, tumor growth rate, and survival time (p<0.001). The HPD-BB4.3 conjugate inhibited the growth of established tumors by more than 40% in comparison with control groups (p<0.05).

GENERATION AND PARTIAL CHARACTERIZATION OF MONOCLONAL ANTIBODIES AGAINST SOLUBILIZED MEMBRANE FRACTION OF HUMAN SPERMATOZOA

Human sperm membrane antigens extracted by deoxycholate (DOC) were used to immunizeBALB/c mice.Hybrid cell lines secreting sperm-specific monoclonal antibodies were generatedby cell fusion in a semi-solid medium and screened by indirect immunofluorescent assay usinglive and methanol-fixed sperm.Out of 850 hybrid clones from cell fusion,28 were shownto secrete sperm-specific antibodies which reacted with the acrosome,equatorial segment,whole surface plasma membrane or tail of spermatozoa.Finally,seven hybrid cell lineswere established and shown to secrete monoclonal antibodies which had no cross-reactivitywith arty human tissues other than testis and sperm.The majority were also shown toinhibit fertilization of mouse oocytes in vitro and human sperm penetration of zona-freehamster ova.Western blot analysis revealed that some of these antibodies reacted withsperm membrane antigens of distinct molecular size.

Clinical correlations of infliximab trough levels and antibodies to infliximab in South Korean patients with Crohn's disease

To investigate the clinical implications of infliximab trough levels (IFX-TLs) and antibodies to infliximab (ATI) levels in Crohn’s disease (CD) patients in Asian countries.METHODSIFX-TL and ATI level were measured using prospectively collected samples obtained with informed consent from CD patients being treated at Asan Medical Center, South Korea. We analyzed the correlations between IFX-TLs/ATI levels and the clinical activity of CD (quiescent vs active disease) based on the CD activity index, C-reactive protein level, and physician’s judgment of patients’ clinical status at enrollment. The impact of concomitant immunomodulators was also investigated.RESULTSThis study enrolled 138 patients with CD (84 with quiescent and 54 with active disease). In patients with quiescent and active diseases, the median IFX-TLs were 1.423 μg/mL and 0.163 μg/mL, respectively (P < 0.001) and the median ATI levels were 8.064 AU/mL and 11.209 AU/mL, respectively (P < 0.001). In the ATI-negative and -positive groups, the median IFX-TLs were 1.415 μg/mL and 0.141 μg/mL, respectively (P < 0.001). In patients with and without concomitant immunomodulator use, there were no differences in IFX-TLs (0.632 μg/mL and 1.150 μg/mL, respectively; P = 0.274) or ATI levels (8.655 AU/mL and 9.017 AU/mL, respectively; P = 0.083).CONCLUSIONIFX-TL/ATI levels were well correlated with the clinical activity in South Korean CD patients. Our findings support the usefulness of IFX-TLs/ATI levels in treating CD patients receiving IFX in clinical practice.

Seeing is believing:anti-PD-1/PD-L1 monoclonal antibodies in action for checkpoint blockade tumor immunotherapy

Structural immunology,focusing on structures of host immune related molecules,enables the immunologists to see what the molecules look like,and more importantly,how they work together.Antibody-based PD-1/PD-L1 blockade therapy has achieved brilliant successes in clinical applications.The recent breakthrough of the complex structures of checkpoint blockade antibodies with their counterparts,pembrolizumab with PD-1 and avelumab with PD-L1,have made it clear how these monoclonal antibodies compete the binding of PD-1/PD-L1 and function to blockade the receptor-ligand interaction.Herein,we summarize the structural findings of these two reports and look into the future for how this information would facilitate the development of more efficient PD-1/PD-L1 targeting antibodies,small molecule drugs,and other protein or non-protein inhibitors.

Pharmacokinetics of monoclonal antibodies ]nd Fc-fusion proteins

There are many factors that can influence the pharma- cokinetics （PK） of a mAb or Fc-fusion molecule with the primary determinant being FcRn-mediatad recycling. Through Fab or Fc engineering, IgG-FcRn interaction can be used to generate a variety of therapeutic anti. bodies with significantly enhanced half-life or ability to remove unwanted antigen from circulation, Glycosyla- tion of a mAb or Fc.fusion protein can have a significant impact on the PK of these molecules, mAb charge can be important and variants with pl values of 1-2 unit difference are likely to impact PK with lower pl values being favorable for a longer half.life. Most mAbs display target mediated drug disposition （TMOO）, which can have significant consequences on the study designs of preclinical and clinical studies. The PK of mAb can also be influenced by anti-drug antibody （ADA） response and off.target binding, which require careful consideration during the discovery stage, mAbs are primarily absor- bed through the lymphatics via convection and can be conveniently administered by the subcutaneous （sc） route in large doses/volumes with co-formulation of hyaluronidase. The human PK of a mAb can be rea- sonably estimated using cynomolgus monkey data and allometric scaling methods.

Effects of anti-CXCR_4 monoclonal antibody 12G5 on proliferation and apoptosis of human acute myelocytic leukemia cell line HL-60

Objective：To investigate the expression of CXCR4 on HL-60 cell line and the proliferation, apoptosis of HL-60 cell line cocultured with bone marrow stromal cells, so as to assess the possibility of 12G5, an anti-CXCR4 monoclonal antibody, in eradicating the minimal residual disease. Methods：The activity of SDF-1 was inhibited by 10 μg/ml 12G5. After treatment with 12G5, the status of adhesion was observed, and the adhesion rates, apoptosis and cell cycles were detected after 24 h of treatment. Cell growth rates were measured by trypan blue exclusion. Cell growth curve was plotted, and the expression of PCNA and apoptosis related protein including PCNA, Bcl-2 and Fas were detected with immunohistochemical technique. Results：（1） There was middling degree expression of CXCR4 on HL-B0 membrane. From 0 h to 6 h, as the time of 12G5 incubation along, the expression of CXCR4 decreased gradually. （2） After treatment for 24 h, the adhesion rates in the experiment group and the control were （39.4±7.9）% and （51.4±5.9）%, respectively. （3）After treatment for 24 h, the percentage of HL-60 cells in G0/G1 phase were （55.21±4.9）%, and that in S phase and G2/M phase were （30.40±4.1）% and （14.39± 5.2）%, respectively, with the corresponding proportions being （44. 67±2.2）%, （45.30±3.7）%, and （10. 03±2.6）% in the control. （4） The percentage of apoptotic HL-60 cells was （8.95±1.7）% in the experiment group, compared to （3. 97±2. 4）% in the control. （5）The survival rates of HL-60 cells decreased markedly at 48 h to 96 h, and the proliferation slowed down at this time duration. （6）The expression of PCNA and Bcl-2 down-regulated significantly, but the Fas protein expression was up-regulated. Conclusion：12G5 could inhibit the capability of adhesion and proliferation of HL-60 cells and it can induce more cells to enter G0/G1 phase and promote apoptosis. It may be helpful by inhibiting the bioactivity of SDF-1 with 12G5 in the therapy of marrow residual disease.

Excellent effects and possible mechanisms of action of a new antibody–drug conjugate against EGFR-positive triple-negative breast cancer

Background:Triple-negative breast cancer(TNBC)is the most aggressive subtype and occurs in approximately 15%–20%of diagnosed breast cancers.TNBC is characterized by its highly metastatic and recurrent features,as well as a lack of specific targets and targeted therapeutics.Epidermal growth factor receptor(EGFR)is highly expressed in a variety of tumors,especially in TNBC.LR004-VC-MMAE is a new EGFR-targeting antibody–drug conjugate produced by our laboratory.This study aimed to evaluate its antitumor activities against EGFR-positive TNBC and further studied its possible mechanism of antitumor action.Methods:LR004-VC-MMAE was prepared by coupling a cytotoxic payload(MMAE)to an anti-EGFR antibody(LR004)via a linker,and the drug-to-antibody ratio(DAR)was analyzed by HIC-HPLC.The gene expression of EGFR in a series of breast cancer cell lines was assessed using a publicly available microarray dataset(GSE41313)and Western blotting.MDA-MB-468 and MDA-MB-231 cells were treated with LR004-VC-MMAE(0,0.0066,0.066,0.66,6.6 nmol/L),and the inhibitory effects of LR004-VC-MMAE on cell proliferation were examined by CCK-8 and colony formation.The migration and invasion capacity of MDA-MB-468 and MDA-MB-231 cells were tested at different LR004-VCMMAE concentrations(2.5 and 5 nmol/L)with wound healing and Transwell invasion assays.Flow cytometric analysis and tumorsphere-forming assays were used to detect the killing effects of LR004-VC-MMAE on cancer stem cells(MDA-MB-468 and MDA-MB-231 cells).The mouse xenograft models were also used to evaluate the antitumor efficacy of LR004-VC-MMAE in vivo.Briefly,BALB/c nude mice were subcutaneously inoculated with MDA-MB-468 or MDAMB-231 cells.Then they were randomly divided into 4 groups(n=6 per group)and treated with PBS,naked LR004(10 mg/kg),LR004-VC-MMAE(10 mg/kg),or doxorubicin,respectively.Tumor sizes and the body weights of mice were measured every 4 d.The effects of LR004-VC-MMAE on apoptosis and cell cycle distribution were analyzed by flow cytometry.Western blotting was used to detect the effects of LR004-VC-MMAE on EGFR,ERK,MEK phosphorylation and tumor stemness marker gene expression.Results:LR004-VC-MMAE with a DAR of 4.02 were obtained.The expression of EGFR was found to be significantly higher in TNBC cells compared with non-TNBC cells(P<0.01).LR004-VC-MMAE inhibited the proliferation of EGFRpositive TNBC cells,and the ICvalues of MDA-MB-468 and MDA-MB-231 cells treated with LR004-VC-MMAE for 72 h were(0.13±0.02)nmol/L and(0.66±0.06)nmol/L,respectively,which were significantly lower than that of cells treated with MMAE[(3.20±0.60)nmol/L,P<0.01,and(6.60±0.50)nmol/L,P<0.001].LR004-VC-MMAE effectively inhibited migration and invasion of MDA-MB-468 and MDA-MB-231 cells.Moreover,LR004-VC-MMAE also killed tumor stem cells in EGFR-positive TNBC cells and impaired their tumorsphere-forming ability.In TNBC xenograft models,LR004-VC-MMAE at 10 mg/kg significantly suppressed tumor growth and achieved complete tumor regression on day 36.Surprisingly,tumor recurrence was not observed until the end of the experiment on day 52.In a mechanistic study,we found that LR004-VC-MMAE significantly induced cell apoptosis and cell cycle arrest at G/M phase in MDAMB-468[(34±5)%vs.(12±2)%,P<0.001]and MDA-MB-231[(27±4)%vs.(18±3)%,P<0.01]cells.LR004-VC-MMAE also inhibited the activation of EGFR signaling and the expression of cancer stemness marker genes such as Oct4,Sox2,KLF4 and EpCAM.Conclusions:LR004-VC-MMAE showed effective antitumor activity by inhibiting the activation of EGFR signaling and the expression of cancer stemness marker genes.It might be a promising therapeutic candidate and provides a potential therapeutic avenue for the treatment of EGFR-positive TNBC.

SPECIFIC UPTAKE OF MONOCLONAL ANTIBODY-CONJUGATED METHOTREXATE BY HUMAN LYMPHOCYTIC LEUKEMIC B CELLS

Objective: To analysis the uptake of free MTX and MTX conjugated to tumor specific monoclonal antibody by target and nontarget cells. Methods: The folate antagonist methotrexate (MTX) was conjugated to two monoclonal antibodies (Mab) directed against human chronic lymphocytic leukemia (CLL), Dal B01 and Dal B02, by an active ester method. Both conjugates were more cytotoxic toward the target tumor cell line D101 than to the nontarget cell line MOLT3, and Dal B02MTX conjugate was more inhibitory to D101 cells than free MTX in a 6 h pulse exposure assay. Results: Drug uptake studies revealed that D101 cells took up much more Dal B01 and Dal B02conjugated MTX than free MTX. The amounts of drug taken up by D101 cells incubated with Dal B01 and Dal B02conjugated MTX were always 3 to 5fold higher than that taken up by MOLT3 cells, although the latter took up more drug when incubated with free MTX. Furthermore, tumor cells incubated with Dal B01 or Dal B02conjugated MTX retained much larger amounts of drug for a prolonged period of time than those incubated with free MTX. Conclusion: The enhanced specific cytotoxicity of Dal B01 and Dal B02MTX conjugates toward target tumor cells is therefore likely due to (I) delivery of larger amounts of MTX to target cells when the drug is conjugated to Mab; (ii) longer retention of Mabconjugated MTX by target cells; and (iii) slow, prolonged release of MTX from the surfacebound or endocytosed conjugates, rendering them into a sustained release dosage form.

LABELING McAb 3H11 AND ITS Fab FRAGMENT WITH ^（211）At AND THEIR IMMUNOREACTIVITIES AND INJURY EFFECTS ON HUMAN GASTRIC CANCER CELLS

An antigastric cancer monoclonal antibody, 3H11, and its Fab fragment, were labeled using p-(211At)-astatobenzoic acid (pAtBA) intermediate, in the yields of more than 30%. The results of in vitro experiments show that 211At-3H11 and 211At-3H11Fab are stable, and have specific immunoreactivities and cytotoxic effects to human gastric cancer cell M85. The cytotoxic effects are dependent on the concentration of 211At-3H11 or 211At-3H11 Fab and obviously stronger than that of Na211At.

Systemic adverse effects and toxicities associated with immunotherapy:A review

Immunotherapy is rapidly evolving secondary to the advent of newer immunotherapeutic agents and increasing approval of the current agents by the United States Food and Drug Administration to treat a wide spectrum of cancers.Immunotherapeutic agents have gained immense popularity due to their tumorspecific action.Immunotherapy is slowly transforming into a separate therapeutic entity,and the fifth pillar of management for cancers alongside surgery,radiotherapy,chemotherapy,and targeted therapy.However,like any therapeutic entity it has its own adverse effects.With the increasing use of immunotherapeutic agents,it is vital for physicians to acquaint themselves with these adverse effects.The aim of this review is to investigate the common systemic adverse effects and toxicities associated with the use of different classes of immunotherapeutic agents.We provide an overview of potential adverse effects and toxicities associated with different classes of immunotherapeutic agents organized by organ systems,as well as an extensive discussion of the current recommendations for treatment and clinical trial data.As we continue to see increasing usage of these agents in clinical practice,it is vital for physicians to familiarize themselves with these effects.

基于高通量表面等离子体共振技术筛选与TPBG具有高亲和力的单克隆抗体研究

目的探讨从制备的单克隆抗体中快速筛选出与滋养层糖蛋白(TPBG)具有高亲和力的单克隆抗体的方法。方法采用高通量表面等离子体共振技术(SPR),将不同种属的TPBG通过氨基偶联的方式固定在GLC传感器芯片上,梯度稀释的抗体作为流通相,筛选能高度亲和TPBG的单克隆抗体,并测定相互作用的动力学常数。抗体1结合人和食蟹猴TPBG,亲和力分别为65.0、31.6 nmol/L;抗体6结合人、小鼠、食蟹猴TPBG,亲和力分别为77.6、92.1、87.7 nmol/L。结果该研究成功筛选出2种能与TPBG特异性结合的单克隆抗体。动力学图谱显示,这2种抗体与TPBG蛋白的特异性结合稳定。结论采用SPR可筛选出与TPBG蛋白具有高亲和力的单克隆抗体,为开发出一些新型的抗肿瘤药物提供一种方法。

阿尔茨海默病的药物治疗新进展

近年来,随着以β淀粉样蛋白(β-Amyloid,Aβ)单抗为代表的疾病修饰治疗(disease-modifying therapies,DMT)药物的出现,阿尔茨海默病(Alzheimer's disease,AD)的药物治疗进入了一个全新的阶段,目前已经有2种靶向淀粉样蛋白的抗体-阿杜卡单抗和伦卡纳单抗相继上市,在全世界引起了广泛关注;同时非Aβ非tau蛋白治疗靶点的药物研发也取得明显进展。本文拟对AD新型药物的研发和应用,包括临床试验阶段和已获批上市的药物治疗进展进行一全面分析和阐述。

生物制剂在甲状腺相关眼病中的研究进展

甲状腺相关眼病(TAO)是一种罕见的器官特异性自身免疫性疾病,发病机制尚未明确。目前治疗仍主要依赖糖皮质激素和传统免疫抑制剂,部分患者对这些药物反应不佳,同时存在治疗相关不良反应,因此迫切需要治疗TAO的新型药物。近年来,随着对TAO发病机制研究的不断深入,针对特定靶点研发的生物制剂层出不穷,其中靶向胰岛素样生长因子-Ⅰ受体(IGF-IR)的替妥木单抗已被美国食品和药物管理局批准用于治疗TAO,还有多种生物制剂处于临床试验阶段。本文通过总结针对IGF-IR、新生儿Fc受体(FcRn)、促甲状腺激素受体(TSHR)、B细胞、细胞因子等的生物制剂在TAO中的临床研究现状,分析其对临床治疗及未来研究趋势的影响,为TAO临床防治和研究提供最新参考。

治疗阿尔茨海默病创新药物研发进展

阿尔茨海默病(AD)是一种中枢神经系统退行性疾病,临床表现为进行性认知功能下降和行为损害,因其发病机制复杂,目前尚无有效的预防和治疗手段。近年来,依据AD发病机制和病理学特点进行靶向治疗逐渐成为新药开发的重点。靶向药物β淀粉样蛋白(Aβ)单克隆抗体阿度奴单抗和来卡内单抗获美国FDA批准用于AD的治疗,调节肠道菌群药物甘露特纳在中国批准上市,β分泌酶抑制剂、抗Aβ疫苗、tau蛋白聚集抑制剂等创新药物也表现出对AD的治疗潜力并先后进入临床试验。本文梳理近年来治疗AD的创新药物研究进展,分析创新药物研发策略,为治疗AD和开展相关新药研究提供参考和思路。

基于QbD理念的单克隆抗体生物类似药研发缺陷管理策略研究

目的为生物类似药的早期研发管理提供参考。方法在阿达木单克隆抗体生物类似药研发、申报实践基础上,结合相关法规、指导原则的要求及审评案例,以药事监管的视角分析单克隆抗体生物类似药研发过程中存在的药学研究缺陷。基于质量源于设计(QbD)理念,从产品质量属性研究全面性、工艺过程控制科学性、外源因子安全性评估充分性、稳定性研究完整性、研究数据规范性五方面提出对应的缺陷管理策略。结果与结论研究缺陷主要表现在药物质量属性研究不全面、工艺过程控制不科学、外源因子安全性评估不充分、稳定性研究不完整、研究数据不规范等方面。在充分研究产品质量属性的前提下,通过产品设计-工艺控制-质量实现实施缺陷管理,提高生物类似药的研发效率及产品研发和申报质量。

阿替利珠单抗不良反应94例文献分析

目的分析阿替利珠单抗发生不良反应(adverse reactions,ADRs)的临床特点与规律,为临床安全用药提供参考。方法搜索中国知网、维普、万方、Web of Science、PubMed数据库,收集关于阿替利珠单抗所致不良反应的报道文献并进行分析,研究时间为2022年4—8月。结果阿替利珠致不良反应报道共94例;其中男性56例(59.57%),女性38例(40.43%),男性占比较高;年龄(62.8±12.0)岁,中老年人居多;多数发生在用药后的90 d内(71例,71.0%);阿替利珠单抗致ADRs累及多个系统/器官,其中以神经系统损害(22例,22.0%)占比最多;3~4级严重ADRs占比最多(64例,64.0%);94例经治疗和(或)停药后,好转或治愈80例,死亡5例。结论阿替利珠单抗所致ADRs涉及不同性别与年龄段病人,累及多个系统/器官,临床使用应随时监测,警惕ADRs的发生,做到及时识别与治疗。

血清IL-17、INF-γ水平与肝细胞癌患者PD-1单抗治疗效果的关系

目的探究血清IL-17、INF-γ水平与肝细胞癌患者PD-1单抗治疗效果的关系。方法选取2023年1月至2023年8月收治的60例肝细胞癌患者作为研究对象,均接受PD-1单抗治疗。比较观察治疗有效患者与无效患者IL-17、INF-γ水平情况,IL-17、INF-γ水平与肝癌细胞患者各临床特征的关系,ROC曲线分析IL-17、INF-γ表达水平对肝癌细胞患者PD-1单抗治疗效果的预测价值关系。结果治疗有效组患者IL-17水平(14.17±2.08)pg/ml低于治疗无效组的(21.28±3.69)pg/ml,INF-γ(37.45±4.52)pg/ml高于对照组的(22.36±3.27)pg/ml(P<0.05)。在临床分期、肝外转移、血管侵犯等方面,分期越高、有肝外转移、有血管侵犯患者的IL-17水平高于分期越低、无肝转移、无血管侵犯患者(P<0.05)。分期越高、有肝外转移、有血管侵犯患者的INF-γ水平低于分期越低、无肝外转移、无血管侵犯患者(P<0.05)。通过ROC曲线的绘制可以得出:(1)IL-17、INF-γ的曲线下面积分别是0.753、0.764,均具有良好的诊断效能,但与2项联合相比,2项联合的曲线下面积0.877均高于各单一指标的曲线下面积(P<0.05);(2)IL-17、INF-γ的cut-off值是6.21 pg/ml、36.52 pg/ml;(3)2项联合的灵敏度(0.935)、特异度(0.952)均明显高于IL-17的(0.552、0.801)和INF-γ的(0.457、0.824)(P<0.05)。结论血清IL-17、INF-γ水平可对细胞癌患者PD-1单抗治疗效果进行预测评估,且两者联合预测的效能更高,具有显著的临床应用价值。

Safety of teplizumab in patients with high-risk for diabetes mellitus type 1:A systematic review

BACKGROUND The incidence of diabetes mellitus type 1(DM1)has been rising worldwide because of improvements in diagnostic techniques and improved access to care in countries with lower socioeconomic status.A new anti-CD4 antibody,Teplizumab,has been shown to delay the progression of DM1 and is the only medication approved for this indication.However,more information is needed about the safety profile of this drug.AIM To identify the odds ratios(OR)of systems-based adverse effects for Teplizumab when compared to Placebo.METHODS An extensive systematic review was conducted from the inception of the medication until December 31,2023.All clinical trials and studies that evaluated Teplizumab vs placebo were included in the initial review.The study protocol was designed using Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines guidelines and was registered in PROSPERO(ID:CRD42024496169).Crude OR were generated using RevMan Software version 5.4.RESULTS After screening and review,5 studies were selected to determine the risk of adverse effects of teplizumab compared to placebo.A total of 561 patients were included in the study population.Total adverse effects and system-based adverse effects were studied and reported.We determined that patients receiving Teplizumab had a higher risk of developing gastrointestinal(GI)(OR=1.60,95%CI:1.01-2.52,P=0.04),dermatological(OR=6.33,95%CI:4.05-9.88,P<0.00001)and hematological adverse effects(OR=19.03,95%CI:11.09-32.66,P<0.00001).These patients were also significantly likely to have active Epstein-Barr Virus infection(OR=3.16,95%CI:1.51-6.64,P<0.002).While our data showed that patients receiving Teplizumab did have a higher incidence of total adverse effects vs placebo,this finding did not reach statistical significance(OR=2.25,95%CI:0.80-6.29,P=0.12).CONCLUSION Our systematic review suggests that Teplizumab patients are at risk for significant adverse effects,primarily related to GI,dermatological,and hematological systems.The total adverse effect data is limited as study populations are small.More studies should be conducted on this medication to better inform the target population of potential adverse effects.

抗体偶联药物在肿瘤治疗中的研究进展

抗体—药物偶联物是由特异性的单克隆抗体和高细胞毒性的化学药物通过连接子组合而成,不仅可以解决单克隆抗体临床效果差、毒副作用较大的问题,还能将化学小分子药物特定地靶向到肿瘤细胞,使药物疗效最大化。目前,随着抗体偶联药物的不断发展,在肿瘤的临床治疗应用也逐渐广泛,具有广阔发展前景。

鸭坦布苏病毒单克隆抗体的制备和治疗效果分析

为了制备、筛选和评价治疗鸭坦布苏病毒(DTMUV)感染的中和抗体,本试验克隆DTMUV E基因,连接至原核表达载体pET-30a(+),转化至大肠杆菌(E.coli)DH5α感受态,提取质粒,鉴定重组质粒pET30-E,转化E.coli BL21感受态,经异丙基-β-D-硫代半乳糖苷(IPTG)诱导后,通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)分析E蛋白表达,使用镍柱亲和层析纯化E蛋白,将E蛋白与弗氏佐剂混合、乳化,免疫6周龄雌性BALB/c小鼠3次后,取小鼠脾脏,分离脾细胞,在聚乙二醇(PEG)的作用下,将脾细胞与SP2/0细胞融合,采用酶联免疫吸附测定(ELISA)筛选分泌抗E蛋白抗体的杂交瘤细胞株,通过Western blot进行单克隆抗体鉴定,通过间接免疫荧光技术检测DTMUV,病毒中和试验评价单克隆抗体中和DTMUV感染的能力,并评价单克隆抗体治疗DTMUV感染小鼠的能力。结果显示,含质粒pET30-E的E.coli BL21经诱导后可在裂解的沉淀物中表达E蛋白;制备的E蛋白单克隆抗体4A1可以与E蛋白发生免疫反应,不与鸭A型肝炎病毒、呼肠孤病毒、新城疫病毒、细小病毒和H9亚型禽流感病毒发生交叉反应;4A1株杂交瘤细胞株分泌的抗体对DTMUV具有中和活性,且能够很好地识别DTMUV并与之发生中和反应,可完全清除感染小鼠血液中的DTMUV。结果表明,本试验制备的单克隆抗体4A1在应对DTMUV感染方面具有潜在治疗价值。