抗人晚期糖基化终产物受体胞外段不同表位单克隆抗体的制备

目的 制备抗人晚期糖基化终产物受体(RAGE)胞外段单克隆抗体。方法 用纯化人重组RAGE胞外段(氨基酸序列23~342)免疫Balb/c小鼠,取其脾细胞与NS-1融合制备杂交瘤,经筛选和两次克隆化,从制备的腹水中纯化单克隆抗体。结果和结论 获得两株杂交瘤细胞B2.2和E10,其分泌单抗的亚型均为IgG2b。经ELISA、Western blot和流式细胞仪鉴定,证明两株单抗均可结合重组RAGE及表达于细胞表面的RAGE,且两株抗体识别的位点为远隔表位,所建立的双夹心法可用于检测可溶性重组RAGE。这两株抗体将为研究RAGE相关疾病提供有效的工具。     

抗小鼠TLR2胞外段单表位抗体TSP-2对C48/80诱导P815细胞脱颗粒的影响

目的观察抗小鼠TLR2胞外段单表位抗体(TSP-2)对C48/80诱导的肥大细胞瘤P815细胞脱颗粒的影响。方法用RT-PCR、免疫组化检测了小鼠肥大细胞瘤P815细胞在mRNA水平的表达及膜表面TLR2与抗体TSP-2的结合;利用激光共聚焦显微镜观察小鼠TLR2胞外段单表位抗体TSP-2对C48/80诱导的肥大细胞瘤P815细胞脱颗粒的影响。结果P815细胞在mRNA水平以及膜表面均有TLR2表达,抗体TSP-2能有效抑制C48/80诱导的肥大细胞瘤P815细胞脱颗粒反应所致的形态学改变与钙流出。结论抗小鼠TLR2胞外段单表位抗体TSP-2可与P815细胞膜表面TLR2结合,并能抑制C48/80诱导的P815肥大瘤细胞脱颗粒。     

博落回总碱对肝癌细胞的毒性作用和体内抗肿瘤作用

目的 观察博落回总碱(TAPP)的体内外抗肿瘤活性作用。方法 MTT法检测TAPP体外对人肝癌Hep3B细胞、小鼠H22肝癌细胞的抑瘤作用;用小鼠移植性肿瘤法检测TAPP对小鼠H22肝癌细胞、S180肿瘤细胞的体内抑瘤作用。测3次重复实验结果。结果 (1)TAPP在体外有明显的细胞毒性作用,可抑制Hep3B、H22细胞增殖,且呈剂量依赖性:其对Hep3B细胞的半数抑制浓度(IC₅₀) 3次重复实验结果分别为3.04、3.98、2.98µg/ml,对H22细胞则分别为2.89、2.21、2.34µg/ml。(2)体内实验表明,TAPP可抑制H22细胞皮下移植瘤的生长,在剂量为每天1、2、4 mg/kg·b.w.时,3次腹腔注射给药实验测得抑瘤率分别为18.6%、35.1%、44.9%,而以每天4 mg/kg·b.w.时口服给药抑瘤率则为7.9%;另外,TAPP可明显延长S180腹水瘤小鼠存活时间,剂量为每天1、2、4 mg/kg·b.w. 时静脉注射给药生命延长率3次重复结果平均分别为24.8%、48.9%、52.7%。结论 TAPP在体内外均有明显的抗肿瘤活性。     

抗小鼠TLR2胞外段单表位抗体TSP-2对OVA诱导的小鼠过敏性哮喘气道炎症的影响

目的观察抗小鼠TLR2胞外段单表位(mTLR2ECD)抗体(TSP-2)对OVA诱导的小鼠过敏性哮喘气道炎症的影响。方法用肺泡灌洗、组织切片及特异染色、免疫组化、ELISA法检测TSP-2对小鼠哮喘模型的气道炎症和炎症细胞的浸润以及肺组织中肺泡及支气管结构的影响。结果发现静脉给予抗体TSP-2能有效抑制气道炎症和炎症细胞的浸润。主要表现在肺泡灌洗液中的白细胞浸润减少、减轻血清中OVA特异性IgE抗体的降低以及抑制OVA诱导的小鼠肺泡毛细血管充血水肿等病理变化。结论抗体TSP-2对过敏性哮喘的病理变化有一定的抑制作用。     

抗小鼠TLR2胞外段单表位抗体TSP-2对脓毒症小鼠肠粘膜核因子-κB和细胞因子表达的影响

目的观察抗小鼠Toll样受体2胞外段(mTLR2ECD)单表位抗体TSP-2对脓毒症小鼠肠粘膜核因子-κB(NF-κB)和细胞因子表达的影响。方法将雄性BALB/c小鼠随机分成4组:假手术组、模型组、TSP-2治疗组和正常兔IgG治疗组。采用盲肠结扎穿孔术制作脓毒症小鼠模型,分别于术后6、12和24 h取回肠组织,HE染色,免疫组织化学方法检测小鼠肠粘膜组织NF-κB的表达,实时荧光定量RT-PCR法检测TNF-α和IL-6 mRNA的表达,ELISA法检测小鼠肠粘膜TNF-α和IL-6蛋白表达水平。结果模型组NF-κB表达水平较假手术组显著上升;TSP-2治疗组NF-κB表达水平较模型组显著下调;模型组各时间点TNF-α和IL-6水平均高于假手术组(P<0.05);TSP-2治疗组与模型组相比,TNF-α和IL-6水平明显降低(P<0.05),TNF-α和IL-6 mRNA水平也低于模型组(P<0.05)。正常兔IgG治疗组与模型组比较,结果无统计学意义(P>0.05)。结论抗体TSP-2能抑制脓毒症小鼠肠道NF-κB的活化,并下调致炎细胞因子TNF-α和IL-6水平,对脓毒血症引起的肠道炎症起到一定的控制作用,并延缓脓毒血症的病情发展。     

新型冠状病毒疫苗免疫BALB/c小鼠后的T细胞表位鉴定

目的 鉴定携带新型冠状病毒S、N和M基因的疫苗在BALB/c小鼠上的T细胞表位。方法 用携带新型冠状病毒S、N和M基因的DNA疫苗和痘病毒载体疫苗免疫6~8周龄雌性BALB/c小鼠,末次免疫1个月内取小鼠脾细胞,用ELISPOT法检测其对新型冠状病毒特异性多肽的T细胞免疫反应,筛选出具有阳性反应的多肽片段,并对其表位进行预测分析。结果 S、N、M基因分别鉴定出12、2、17条阳性多肽。S基因中多肽S49、S50、S100、S101、S102、S103阳性反应率为100%,且免疫刺激指数最高;覆盖M蛋白全长的41条多肽中有7条能刺激50%以上的小鼠产生阳性反应。预测分析结果显示,S、N、M基因分别有10、1、16个MHC-1 H-2限制性表位,主要位于S基因的RBD区、N基因RBD区以及M基因的跨膜区。这些表位在SARS-CoV-2及其变异株、SARS-CoV中高度保守。结论 新型冠状病毒DNA疫苗和痘病毒载体疫苗诱导了针对多个基因、多个表位的特异性T细胞应答,未来通用型疫苗设计应包含能诱导细胞免疫的多个抗原,特别是包含保守表位的结构抗原。     

TSP-2对小鼠溃疡性结肠炎NF-κB DNA结合活性和表达的影响

目的研究抗小鼠Toll样受体2（Toll-like receptor 2）胞外段（mTLR2ECD）单表位抗体TSP-2对溃疡性结肠炎小鼠肠粘膜
核因子NF-κB DNA结合活性和表达的影响。方法将实验小鼠随机分为4组：正常对照组、模型组、TSP-2治疗组和兔IgG组,其
中模型组、TSP-2治疗组和兔IgG组均给予5%葡聚糖硫酸钠7 d以制备溃疡性结肠炎小鼠模型,停用造模药后TSP-2治疗组和
兔IgG组给予后续干预治疗7 d。计算溃疡性结肠炎疾病活动指数（DAI）,14 d后处死小鼠,取结肠行HE染色;利用ELISA法测
小鼠肠粘膜NF-κB P65 DNA结合活性;用Western blot 法测定NF-κB p65蛋白水平,并进行统计学分析。结果模型组NF-κB
p65 DNA结合活性明显高于对照组（P<0.05）;模型组NF-κB P65 蛋白表达明显高于NC组（P<0.05）;TSP-2治疗组小鼠第10~
14 天DAI明显低于模型组（P<0.05）;TSP-2 治疗组NF-κB p65 DNA结合活性明显低于MD组（P<0.05）;TSP-2 治疗组NF-κB
p65蛋白表达水平显著低于模型组（P<0.05）;兔IgG组与模型组比较,实验结果无统计学意义（P>0.05）。结论TSP-2可以明显
抑制NF-κB表达和活性,通过调节肠道过度的免疫反应和抗炎作用,在溃疡性结肠炎治疗中发挥作用。
     

抗人晚期糖基化终产物受体胞外段不同表位单克隆抗体的制备

目的 制备抗人晚期糖基化终产物受体(RAGE)胞外段单克隆抗体。方法 用纯化人重组RAGE胞外段(氨基酸序列23～342)免疫Balb／c小鼠，取其脾细胞与NS—1融合制备杂交瘤，经筛选和两次克隆化，从制备的腹水中纯化单克隆抗体。结果和结论 获得两株杂交瘤细胞B2．2和E10，其分泌单抗的亚型均为IgG2b。经ELISA、Westernblot和流式细胞仪鉴定，证明两株单抗均可结合重组RAGE及表达于细胞表面的RAGE，且两株抗体识别的位点为远隔表位，所建立的双夹心法可用于检测可溶性重组RAGE。这两株抗体将为研究RAGE相关疾病提供有效的工具。     

抗人血红蛋白单克隆抗体的制备及鉴定

目的 运用杂交瘤技术制备抗血红蛋白(hemoglobin, Hb)单克隆抗体,为进一步探讨Hb在帕金森病中的发病机制及早期诊断中的意义奠定基础。方法 用重组人Hb作为抗原免疫Balb/c小鼠,并将其脾细胞与同系骨髓瘤细胞融合制备杂交瘤,经3次有限稀释法及酶联免疫吸附试验法筛选获得抗人Hb单克隆抗体杂交瘤株5B1。腹腔注射于免疫抑制Balb/c小鼠产生腹水,并利用Dot blotting、酶联免疫吸附实验,免疫共沉淀、Western blotting法及免疫组织化学染色法进一步测定抗体效价,获得一特异性抗Hb单克隆抗体。结果 成功制备出1株高亲和力、特异性好的抗Hb单克隆抗体。结论 制备的单克隆抗体对Hb具有特异性(包括人血、猴及小鼠),为进一步研究Hb在帕金森病(Parkinson's disease,PD)中的作用提供了有力的特异性工具。     

抗人ζ珠蛋白链不同表位单克隆抗体的制备和鉴定

目的 制备和鉴定抗ζ珠蛋白链单克隆抗体。方法 用纯化的重组ζ珠蛋白链免疫Balb/c小鼠,取其脾细胞与NS-1细胞融合制备杂交瘤,经重组的ζ珠蛋白链筛选和3次克隆化,从制备的腹水中纯化单克隆抗体。结果和结论 获得3株杂交瘤细胞1A12、3H9及4D11,其分泌单抗的亚型分别为IgG2a、IgG1和IgG1;双抗体夹心ELISA结果证明3株单抗均可结合重组的及东南亚(--^SEA)缺失型地中海贫血基因携带者血中的ζ珠蛋白链,且其中1A12和3H9识别的位点为不同表位。这三株抗体将为筛选地中海贫血基因携带者的免疫检测提供有力工具。     

抗小鼠TLR-2胞外段B细胞识别表位抗体抑制肉瘤生长的初步研究

OBJECTIVE: To observe the effect of the antibody to a B cell epitope on mouse Toll-like receptor-2 (mTLR-2) extracellular domain on the growth of murine fibrosarcoma. METHODS: An immunogen was prepared by conjugating a 20-mer peptide, synthesized on the basis of prediction for B cell dominant epitope of mTLR-2, to a carrier protein. Rabbit polyclonal antibodies against B cell epitope of mTLR-2 were prepared and purified, and characterized by Western blotting, immunohistochemistry and flow cytometry. Murine fibrosarcoma S180 cells were inoculated into the left hindlimbs of Swiss mice, and in the treatment group, the injected cells were mixed with 100 microg anti-mTLR-2 antibody TSP-1, with the same dose of irrelevant rabbit IgG in the IgG control group and with buffer solution only in the blank control group. The mice in each group were anesthetized and sacrificed on days 10, 14, and 18 following the injections, respectively, and the weight of the tumors was measured and the inhibition rate calculated. RESULTS: A distinct band for the antibody TSP-1 was shown at the relative molecular mass of 95 000 by Western blotting. The antibody TSP-1 could also react with native mTLR-2 expressed on J774A.1 cells, as identified by immunohistochemistry and flow cytometry. The growth of the fibrosarcoma S180 was obviously inhibited by injections with the antibody TSP-1 mixture, and the weight of tumor in mice treated with TSP-1 was significantly lower than that in the two control groups (P<0.05), with an inhibition rate of 77% on day 10, 51% on day 14 and 53% on day 18. CONCLUSION: Local application of the antibody against B cell epitope on mTLR-2 extracellular domain can inhibit the growth of murine fibrosarcoma, especially in the early stage of tumor proliferation.     

抗小鼠TLR2胞外段单表位抗体TSP-2对酵母多糖诱发小鼠腹膜炎的影响

目的 观察抗小鼠TLR2胞外段(mTLR2ECD)单表位抗体TSP-2对酵母多糖诱发的小鼠腹膜炎的主要作用.方法 用腹腔灌洗、细胞计数及特异染色、免疫组化、ELISA 等各种方法检测了抗体TSP-2对模型动物的扭身次数、腹腔灌洗液的伊文思兰渗出、腹腔白细胞浸润的数量变化、腹腔肥大细胞脱颗粒情况以及腹腔灌洗液离心上清中PAF和TNFa水平的影响.结果 与PBS处理组和正常兔IgG组相比,抗体TSP-2能促使小鼠因腹腔炎症引起的扭身次数减少,腹腔灌洗液中伊文思蓝OD值降低,腹腔灌洗液中白细胞渗透减少以及C48/80诱导的腹腔肥大细胞脱颗粒降低.结论 抗体TSP-2可能通过对肥大细胞脱颗粒的抑制作用减轻酵母多糖诱导的小鼠腹膜炎炎症症状.

Abstract：

Objective To observe the effect of the antibody TSP-2 against a single epitope of mouse Toll-like receptor 2 extracellular domain (mTLR2ECD) on the inflammation in mice with zymosan A-induced peritonitis. Methods In mice with peritonitis induced by intraperitoneal injection of zymosan A, pretreatments with PBS, normal rabbit IgG and TSP-2 antibody at two different doses (2.5 and 5.0 mg/kg) were administered via the tail vein. Six hours after intraperitoneal injection of zymosan A, Evans blue was injected through the tail vein, and the frequency of writhing of the mice within 20 min were recorded. The mice were then sacrificed for peritoneal lavage, and the lavage fluid was collected to assess the exudation of Evans blue in the supernatant. The peritoneal leukocyte count, mast cell degranulation and release of such inflammatory mediators as platelet activating factor (PAF) and tumor necrosis factor-a (TNFa) in the lavage fluid were observed by cell counting, specific cell staining, immunohistochemistry and enzyme-linked immunosorbent assay (ELISA). Results Compared with PBS or rabbit IgG groups, TSP-2 treatment resulted in significantly reduced writhing response of the mice and lowered Evans blue exudation and leukocyte count in the peritoneal lavage, with also decreased degranulation of the mast cells induced by C48/80. Conclusion TSP-2 antibody against a single epitope of mTLR2ECD inhibits the inflammatory response in mice with zymosan A-induced peritonitis.     

血小板反应蛋白-1在氧诱导小鼠视网膜病变模型中的表达及意义

目的: 检测血小板反应蛋白-1 (thrombonspondin-1,TSP-1) 在氧诱导视网膜病变 (oxygen-induced retinopathy,OIR) 模型小鼠视网膜中的表达,探讨其在视网膜新生血管中的作用。方法: 随机选取7日龄C57BL/6J新生小鼠40只,分为模型组(n=20)及正常对照组(n=20)。模型组小鼠通过高氧诱导的方法建立OIR模型。于小鼠出生后第7,9,11天时两组各随机抽取5只小鼠,取视网膜组织采用RT-PCR法检测TSP-1 mRNA的表达水平; 并于小鼠出生后第11天时两组随机取5只小鼠,运用荧光造影视网膜铺片对视网膜新生血管进行形态学观察。结果: 出生后第11天时,正常组小鼠视网膜铺片显示视网膜血管分布呈均匀的网状结构,而模型组小鼠视盘周围可见大片无灌注区,视网膜大血管扩张,仅在周边见少量毛细血管分布,为典型的OIR早期表现。出生后第7天,模型组与对照组小鼠视网膜组织中TSP-1 mRNA表达水平差异无统计学意义(P>0.05); 出生后第9天,模型组小鼠视网膜组织中TSP-1 mRNA表达水平下降(P<0.05); 出生后第11天模型组小鼠视网膜组织中TSP-1 mRNA表达水平明显低于正常组(P<0.01),且较第9天模型组亦下降(P<0.05)。结论: 在新生小鼠OIR模型视网膜血管生长发育抑制期,视网膜组织中TSP-1 mRNA的表达逐渐下降,提示TSP-1可能作为负调节因子在早期参与视网膜新生血管的形成过程。     

Effects of the configuration of a multi-epitope chimeric malaria DNA vaccine on its antigenicity to mice

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Protection of erythropoietin on experimental spinal cord injury by reducing the expression of thrombospondin-1 and transforming growth factor-β

Background Erythropoietin （EPO） functions as a tissue-protective cytokine in addition to its crucial hormonal role in red cell production and neuron protection. This study aimed to determine the neuron protective effect of erythropoietin on experimental rats enduring spinal cord injury （SCI） by assessing thrombospondin-1 （TSP-1） level and transforming growth factor-β （TGF-β） in the development of a rat model of SCI. Methods Sixty Sprague-Dawley rats were randomly assigned to three groups： sham operation control group, SCI group and EPO treatment group. By using a weight-drop contusion SCI model, the rats in the SCI group and EPO treatment group were sacrificed at 24 hours and 7 days subsequently. The Basso, Beattie, and Bresnahan （BBB） scores were examined for locomotor function. Pathological changes were observed after HE staining. The expressions of thrombospondin-2 （TSP-1） and TGF-β were determined by immunohistochemical staining and Western blotting. Results Slighter locomotor dysfunction was discovered and it was recovered abruptly as higher BBB scores were found in the EPO treatment group than in the SCI group （P 〈0.01）. Pathologically, progressive disruption of the dorsal white matter and regeneration of a few neurons were also observed in SCI rats. TSP-1 and TGF-β expression increased at 24 hours and 7 days after SCI in the injured segment, and it was higher in the SCI group than in the EPO treatment group. Spinal cord samples from the animals demonstrated a TSP-1 optical density of 112.2±6.8 and TSP-1 positive cells of 5.7±1.3 respectively. After injury, the TSP-1 optical density and cell number increased to 287.2±14.3/mm^2 and 23.2±2.6/mm^2 at 24 hours and to 232.1±13.2/mm^2 and 15.2±2.3/mm^2 at 7 days respectively. When EPO treated rats compared with the SCI rats, the TSP-1 optical density and cell number decreased to 213.1 ±11.6/mm^2 and 11.9±1.6/mm^2 at 24 hours and to 189.9±10.5/mm^2 and 9.3±1.5/mm^2 at 7 days, respectively （P 〈0.01）. In the SCI rats, the TGF-β optical density and positive neuron number were 291.4±15.2/mm^2 and 28.8±4.9/mm^2 at 24 hours and 259.1±12.3/mm^2 and 23.9±4.1/mm^2 at 7 days respectively. They decreased in the EPO treated rats to 222.8±11.9/mm^2 and 13.7±2.1/mm^2 at 24 hours and to 196.5±9.7/mm^2 and 8.7±2.2/mm^2 at 7 days （P 〈0.01）. Conclusions Increased expression of TSP-1 and TGF-β can be found in the injured segment of the spinal cord at 24 hours and 7 days after injury. EPO treatment can effectively prevent pathological alterations from severe spinal cord injury by reduced expression of TSP-1 and TGF-β.     

抗β2GPI抗体促进巨噬细胞摄取oxLDL加速动脉粥样硬化进展

目的：研究抗β2糖蛋白Ⅰ(β2GPⅠ)抗体在apoE基因敲除(apoE^-/-)小鼠巨噬细胞摄取功能及动脉粥样硬化之间的关系。方法：将雄性apoE^-/-小鼠随机分为抗β2GPI抗体组和模型组,每组8只,均给予高脂饲料喂养8周,其中,抗β2GPI抗体组小鼠腹腔注射抗β2GPI抗体,模型组小鼠腹腔注射同等剂量0.9%氯化钠溶液稀释的同源对照IgG抗体。8周后解剖小鼠,分离小鼠主动脉根组织,行Movat染色、免疫组织化学染色,观察主动脉根部动脉粥样硬化斑块的病理组织学特点;Western blotting检测主动脉斑块中内质网应激和巨噬细胞的标志蛋白表达;腹腔巨噬细胞体外培养,抗β2GPI抗体组和模型组分别加入氧化型低密度脂蛋白(oxLDL)+抗β2GPI抗体、oxLDL刺激24 h,观察巨噬细胞摄取oxLDL情况。结果：Movat染色显示,抗β2GPI抗体组小鼠主动脉斑块增多,斑块面积较大,胶原纤维、平滑肌纤维含量均减少,与模型组相比差异有统计学意义(P<0.05～P<0.01)。免疫组织化学染色显示,抗β2GPI抗体组小鼠主动脉根部斑...     

抗FasL抗体在小鼠病毒性心肌炎治疗中的作用

目的：探讨抗FasL抗体对小鼠柯萨奇B3（CVB3）病毒性心肌炎心肌细胞凋亡及病毒滴度的调节作用。方法：随机将80只Balb／c小鼠分为空白对照组、病毒对照组、IgG对照组及抗FasL抗体治疗组,于感染后第10天每组处死8只并取其心脏,光镜检查的心肌病理变化,原位末端标记法（TUNEL）检测小鼠心肌组织的细胞凋亡,用空斑形成技术（PFU法）测定心肌病毒滴度。结果：抗FasL抗体治疗组小鼠死亡率、心肌病变积分、心肌细胞凋亡率、心肌病毒滴度均明显低于病毒对照组及IgG对照组（P均〈0．01）。结论：抗FasL抗体可降低小鼠CVB,病毒性心肌炎心肌内病毒滴度、减少心肌细胞凋亡和心肌损伤．     

Anti-angiogeneic Target Therapy for Cancer with Vaccine Based on the Recombinant Chicken FGFR-1 in Tumor-bearing Mice

To explore the anti-tumor effect of immunotherapy with recombinant protein vaccine based on FGFR-1 of chicken (cFR-1) in a mouse Meth A fibrosarcoma model, tumor volume and sur- vival rate of the mice were observed at a 3-day interval. Microvessel density (MVD) was detected by immunohistochemistry. Auto-antibodies against self-FGFR-1 were detected by Western blotting and ELISA, respectively. The anti-FGFR-1 antibody-producing B cells (APBCs) were detected by enzyme-linked immunospot (ELISPOT) assay. Eighteen days after inoculation of tumor cells, the tumor volume was significantly smaller in cFR-1-immunized group than in mouse FGFR-1 (mFR-1) immunized group and normal saline (NS) control group (P<0.05), and the survival time was signifi- cantly longer in cFR-1-immunized group than in the control groups (P<0.01). MVD was significantly lower in cFR-1-immunized group than in mFR-1-immunized group and NS group (16.8 ±5.6 vs 64.6±1.8 and 59.6±8.7, P<0.01). Antibodies against self-FGFR-1 were found in mFR-1-immunized group, the major antibody subclasses were IgG1 and IgG2b. Compared with the two control groups, the numbers of APBCs in cFR-1-immunized group were significantly increased (P<0.01) These re- sults demonstrated that the cFR-1-related anti-angiogenesis protein vaccine could induce the produc- tion of auto-antibodies against self-FGFR-1, which futher inhibit angiogenesis and growth of solid tumor.     

小鼠TLR-2N末端基因的克隆表达和抗体制备

目的 克隆mTLR-2基因氨基端序列,并表达和纯化N端融合蛋白,制备抗mTLR-2 N末端的多克隆抗体.方法 采用PCR技术扩增编码mTLR-2N端1～153氨基酸的基因,并将其克隆到pET32A载体中,测序验证;用大肠杆菌表达融合蛋白,并以Probond树脂柱纯化;免疫家兔制备多克隆抗体,采用免疫组化、流式细胞术及间接ELISA检测抗体特异性及效价.结果 成功构建了融合表达载体pET-N,表达和纯化了相应的融合蛋白,所制备多克隆抗体可与pGEX-N表达的融合蛋白反应,并可结合表达TLR-2的RAW264.7及转染mTLR-2全长基因的CHO细胞.结论 获得了mTLR-2 N末端重组融合蛋白及其可与天然mTLR-2结合的多克隆抗体.