CN117343173A - 人IL-1β蛋白结合分子及其编码基因和应用 - Google Patents
人IL-1β蛋白结合分子及其编码基因和应用 Download PDFInfo
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- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
本发明公开了一种人IL‑1β蛋白结合分子及其编码基因和应用。该人IL‑1β蛋白结合分子具有与人IL‑1β蛋白特异结合的抗原结合位点，抗原结合位点含有至少一个重链可变区和至少一个轻链可变区，所述的轻链可变区的氨基酸序列如SEQ ID No.44所示，所述的重链可变区的氨基酸序列如SEQ ID No.45所示。本发明的人IL‑1β蛋白结合分子具有高亲和力和高阻断活性，具有很大的疾病治疗价值。
Description
本申请是申请号为：202210438106.4、分案提交日为2022年4月25日、申请日为：2019年3月7日、发明名称为人IL-1β蛋白结合分子及其编码基因和应用的中国发明专利申请的分案申请。
技术领域
本发明属于生物制药技术领域，具体涉及一种人IL-1β蛋白结合分子及其编码基因和应用。
背景技术
IL-1β是IL-1家族众多成员中研究的最为深入的成员。包括免疫细胞(如单核细胞和巨噬细胞)在内的很多细胞都可以产生并分泌。IL-1β的释放大致分为3个步骤：i)无活性前体的产生；ii)前体的成熟活化；iii)活化蛋白向胞外的分泌。IL-1β初始产生时是一个31KD的前体，接着裂解活化产生17KD的成熟形式，这样才可以与IL-1受体结合。Caspase-1是主要的裂解蛋白，其可以在IL-1β上两个位点进行切割(D26位和D116位)，从而得到26KD的前蛋白和17KD的成熟形式蛋白。此外，其他的一些蛋白酶如PR-3，Neutrophil elastase等也能切割IL-1β(Andrei C,Margiocco P,Poggi A,et al.Phospholipases Cand A2control lysosome-mediated IL-1βsecretion:Implications for inflammatoryprocesses[J].Proceedings of the National Academy of Sciences of the UnitedStates of America,2004,101(26):9745-9750；Van Damme J,De Ley M,Opdenakker G,etal.Homogeneous interferon-inducing 22K factor is related to endogenouspyrogen and interleukin-1.[J].Nature,1985,314(6008):266-268；Zsebo K M,WypychJ,Yuschenkoff V N,et al.Effects of Hematopoietin-1and Interleukin 1Activitieson Early Hematopoietic Cells of the Bone Marrow[J].Blood,1988,71(4):962-968)。
IL-1β具有很强的生物学活性，包括：(1)介导炎症反应：IL-1β不仅本身能够引起炎症反应，而且可以诱导环氧化酶2(cyclooxygenase-2,COX-2)、iNOS、IL-6等其他炎症因子的表达，从而进一步激活基质细胞和免疫细胞产生更多的IL-1β参与炎症反应；(2)免疫调节作用，协同刺激I细胞，诱导许多细胞产生其他淋巴因子；(3)参与恶液质的形成，有致负氮平衡的效应，可以刺激骨骼肌分解蛋白质；(4)诱导急性时相蛋白，参与急性期反应；(5)诱导成纤维细胞增殖等。
白介素IL-1β释放是早期炎症反应的关键因子，IL-1β与受体IL-1RI结合后与辅助受体IL-1RAcP作用形成IL-1beta/IL-1RI/IL-1RAcP三元复合物，激活靶细胞内的NF-κB和MAPKs信号通路，从而诱导一系列炎症相关分子的表达。有大量的基础和临床数据显示IL-1分泌在急性痛风中起关键作用，通过与细胞因子或其受体结合阻断IL-1的治疗是减少炎症风暴的策略。抑制IL-1分泌是可行的，并且已经有许多IL-1抑制剂可用，因此它可以补充可用的方法来缓解急性痛风发作(So A,Dumusc A,Nasi S,et al.The role of IL-1ingout:from bench to bedside[J].Rheumatology,2018)。
慢阻肺患者气道中IL-1β和系统炎症与频繁急性加重相关，并可能通过恶性循环来介导既往和未来的急性加重。(Fu J,Mcdonald V M,Baines K J,et al.Airway IL-1βand Systemic Inflammation as Predictors of Future Exacerbation Risk in Asthmaand COPD[J].Chest,2015,148(3):618-629)。此外，IL-1β通过促进癌症干细胞和上皮细胞间质转化来支持转移。IL-1β还可以参与Th17分化，参与Th17相关细胞因子的产生。(Tominaga K,Yoshimoto T,Torigoe K,et al.IL-12synergizes with IL-18or IL-1βforIFN-γproduction from human T cells[J].International Immunology,2000,12(2):151-160)。
IL-1β在慢性局部炎症过度表达，也会促进肿瘤发生转化。IL-1β在癌症发生的晚期具有重要作用，可以在肿瘤微环境中产生，并通过与血管生成因子(如VEGF)的细胞信号传导相互作用驱动血管生成，实现肿瘤转移扩散(Voronov E,Carmi Y,Apte R N,etal.The role IL-1in tumor-mediated angiogenesis[J].Frontiers in Physiology,2014:114-114)。此外，吸入肺部的硅石或石棉等异物可通过激活白介素-1β等促炎症细胞因子引起肺癌，而且IL-1β可以可诱导肿瘤细胞分泌内源性TNF-α促进肿瘤生长。因此，抑制IL-1β表达也可以达到预防和治疗肿瘤的作用。
阻断炎症因子的释放可以比较有效的进行抗炎症治疗。针对IL-1β细胞因子，抗IL-1β抗体可以阻断IL-1β与其受体的结合，阻断下游的信号传导。诺华公司的卡纳单抗(Canakinumab，商品名Ilaris)是一种全人源高亲和力抗IL-1β抗体，最初卡纳单抗被批准用于2岁及以上儿童幼年特发性关节炎治疗。基于两项国际随机、安慰剂对照试验，显示了卡纳单抗对幼年特发性关节炎的疗效。在试验1中，单次注射卡纳单抗后，33％患者在15天内出现无疾病活动。试验2证实了这些结果，在治疗2年后，82％的患者具有持续疗效(Vanoni F,Minoia F,Malattia C,et al.Biologics in juvenile idiopathicarthritis:a narrative review.[J].European Journal of Pediatrics,2017,176(9):1147-1153)。随着研究的深入，卡纳单抗在痛风、动脉粥样硬化、冠心病、肺癌等疾病领域都取得了临床试验的成功。
从新型的治疗性抗体分子、提高药物疗效、降低病人使用成本等方面考量，开发新的抗IL-1β抗体仍然有其必要性。
发明内容
本申请的发明目的是提供一种新型的人IL-1β蛋白结合分子。
为实现上述发明目的，本申请的技术方案如下：
一种人IL-1β蛋白结合分子，所述的人IL-1β蛋白结合分子具有与人IL-1β蛋白特异结合的抗原结合位点，该抗原结合位点含有至少一个重链可变区和至少一个轻链可变区，所述的重链可变区具有三个重链互补决定区，三个重链互补决定区的氨基酸序列依次如SEQ ID No.11、SEQ ID No.12和SEQ ID No.13所示；所述的轻链可变区具有三个轻链互补决定区，三个轻链可变区的氨基酸序列依次如SEQ ID No.28、SEQ ID No.29和SEQ IDNo.30所示。
本申请以市售的重组人IL-1β蛋白(novoprotein，CG93)作为抗原免疫小鼠，取免疫小鼠的脾细胞与小鼠骨髓瘤细胞融合制备杂交瘤细胞以表达鼠源人IL-1β多抗，从鼠源人IL-1β多抗中筛选具有优良阻断功能的鼠源人IL-1β单抗，然后根据鼠源人IL-1β单抗的重链可变区和轻链可变区氨基酸序列，结合人抗体恒定区序列设计人鼠嵌合抗体，经人源化改造后获得了本申请的人IL-1β蛋白结合分子，该人IL-1β蛋白结合分子能够特异性结合人IL-1β蛋白，阻断IL-6的分泌水平，具有与卡纳单抗(Canakinumab，商品名Ilaris)相当的功能抑制活性，而与全人源化的卡纳单抗相比，本申请的人IL-1β蛋白结合分子是一个全新序列的抗体分子，具有高亲和力和高阻断活性，具有很大的疾病治疗价值。
在上述的人IL-1β蛋白结合分子中，重链可变区的重链框架区以及轻链可变区的轻链框架区均来自于人的抗体胚系基因序列或与人的抗体胚系基因序列具有至少90％同源性的基因序列。
作为优选，在上述的人IL-1β蛋白结合分子中，所述的重链可变区具有四个重链框架区，四个重链框架区的氨基酸序列依次如SEQ ID No.7、SEQ ID No.8、SEQ ID No.9和SEQID No.10所示。
作为优选，在上述的人IL-1β蛋白结合分子中，所述的轻链可变区具有四个轻链框架区，四个轻链框架区的氨基酸序列依次如SEQ ID No.24、SEQ IDNo.25、SEQ ID No.26和SEQ ID No.27所示。
综合而言，上述的人IL-1β蛋白结合分子具有如SEQ ID No.40所示的重链可变区和如SEQ ID No.41所示的轻链可变区。
此外，本发明还提供了另外两种人IL-1β蛋白结合分子，这两种人IL-1β蛋白结合分子的开发方式与上述人IL-1β蛋白结合分子相同，仅氨基酸序列不同。其中一种人IL-1β蛋白结合分子中，所述的轻链可变区的氨基酸序列如SEQ IDNo.42所示，重链可变区的氨基酸序列如SEQ ID No.43所示。
另一种人IL-1β蛋白结合分子中，所述的轻链可变区的氨基酸序列如SEQ IDNo.44所示，重链可变区的氨基酸序列如SEQ ID No.45所示。
本申请的人IL-1β蛋白结合分子可以是人源化的嵌合抗体，也可以是抗原结合片段或单链抗体可变区片段。其中，嵌合抗体的抗体亚型优选为IgG1、IgG2或IgG4。
嵌合抗体和抗原结合片段除了具有重链可变区和轻链可变区外，还具有重链恒定区和轻链恒定区(氨基酸序列如SEQ ID No.35所示)，区别在于：抗原结合片段仅具有重链恒定区1，且重链恒定区1与轻链恒定区通过链间二硫键相连；而嵌合抗体则具有完整的重链恒定区：重链恒定区1-铰链区-重链恒定区2-重链恒定区3(氨基酸序列如SEQ ID No.34所示)；而在单链抗体可变区片段中，重链可变区和轻链可变区直接通过-(GGGGS)3-短肽相连。
本申请还提供了编码所述的人IL-1β蛋白结合分子的核苷酸分子，以及含有所述的核苷酸分子的表达载体和重组细胞，该重组细胞可以是原核细胞或真核细胞，如CHO细胞、293细胞、大肠杆菌细胞和酵母细胞等，在构件不同的重组细胞时，可以根据不同细胞的密码子偏好对所述的人IL-1β蛋白结合分子的密码子进行优化，进而获得相应的核苷酸分子。
本申请还提供了所述的人IL-1β蛋白结合分子在制备IL-1介导型疾病治疗药物中的应用，其中，所述的IL-1介导型疾病包括自身免疫相关疾病，如幼年特发性关节炎、痛风、哮喘、免疫性脑脊髓炎、炎症性肠病、银屑病、白癜风、糖尿病、动脉粥样硬化、系统性红斑狼疮、硬皮病、皮肌炎、胰腺炎、肾炎、慢性阻碍性肺病、肺纤维化；以及癌症，如胃癌、肝癌、胰腺癌、结肠癌、直肠癌、肺癌、膀胱癌，前列腺癌、宫颈癌、卵巢癌、输卵管癌、乳腺癌、白血病、淋巴瘤、骨髓瘤、神经胶质瘤和骨肉瘤。
上述的IL-1介导型疾病治疗药物可以单独使用也可以与其他对症药物联用，优选以注射制剂的形式使用。
本申请还提供了所述的人IL-1β蛋白结合分子在制备双特异性抗体中的应用，在用于制备双特异性抗体时，所述的人IL-1β蛋白结合分子应以人源化的抗原结合片段或单链抗体可变区片段的形式作为双特异性抗体的其中一个结合臂。
与现有技术相比，本申请的有益效果体现在：
本申请以市售的重组人IL-1β蛋白(novoprotein，CG93)作为抗原免疫小鼠，取免疫小鼠的脾细胞与小鼠骨髓瘤细胞融合制备杂交瘤细胞以表达鼠源人IL-1β多抗，从鼠源人IL-1β多抗中筛选具有优良阻断功能的鼠源人IL-1β单抗，然后根据鼠源人IL-1β单抗的重链可变区和轻链可变区氨基酸序列，结合人抗体恒定区序列设计人鼠嵌合抗体，经人源化改造后获得了本申请的人IL-1β蛋白结合分子，该人IL-1β蛋白结合分子能够特异性结合人IL-1β蛋白，阻断IL-6的分泌水平，具有与卡纳单抗(Canakinumab，商品名Ilaris)相当的功能抑制活性；本申请的抗人IL-1β蛋白抗体是新型的抗体分子序列，具有高亲和力和高阻断活性，具有很大的疾病治疗价值。
附图说明
图1为各亚克隆杂交瘤细胞培养上清抑制IL6因子释放实验结果一；
图2为各亚克隆杂交瘤细胞培养上清抑制IL6因子释放实验结果二；
图1和图2中，“Sample”表示亚克隆杂交瘤细胞培养上清样本，“inhibition％”表示亚克隆杂交瘤细胞培养上清抑制IL6因子释放的抑制率(％)；
图3为抗IL-1β人鼠嵌合抗体抑制IL6因子释放的IC50测定结果；
图3中，“Ab conc.(nM)”表示抗体浓度(nM)，“IL-6(pg/ml)”表示IL6的分泌水平(pg/ml)，A13-8、A13-60、B8-8、B27-4、C20-2表示不同抗IL-1β人鼠嵌合抗体，CAN和IgG1分别为阳性对照抗体和阴性对照抗体。
具体实施方式
下面结合附图和具体实施方式对本发明的技术方案做进一步详细说明。
实施例1制备IL-1β鼠单抗
为了生成针对IL-1β的鼠抗，用市售的重组人IL-1β(novoprotein，CG93)作为抗原，免疫BALB/c小鼠，然后通过ELISA筛选血浆效价，筛选出具有抗重组人IL-1β免疫球蛋白最高滴度的小鼠进行杂交瘤融合。具体步骤如下：
(1)免疫小鼠
首先，每只小鼠使用50μg重组人IL-1β皮下免疫BALB/c小鼠(共5只)，然后，交替使用Gold Adjuvant liquid(Sigma,T2684)和/>Alum Adjuvant(Thermo,77161)皮下免疫BALB/c小鼠(共免疫5次)。
(2)血浆效价检测
将免疫小鼠剪尾取血，通过ELISA筛选血浆效价，以对各免疫小鼠的免疫应答进行检测，筛选出具有抗重组人IL-1β免疫球蛋白最高滴度的小鼠。
用1μg/ml的重组人IL-1β包被ELISA板，每孔100μl于4℃过夜孵育；用200μl/孔PBS/Tween(0.1％)洗板，随后用200μl/孔PBS/Tween(0.1％)中的3％牛血清白蛋白进行封闭1小时；洗板后，每孔加入来自重组人IL-1β免疫小鼠血清稀释液，37℃孵育2小时，洗板；然后用稀释后的带HRP山羊抗小鼠IgG(H+L)(北京博奥龙免疫技术有限公司,BF03001)抗体37℃孵育1小时；洗涤后，每孔用100μl TMB显色液(Biopanda,TMB-S-002)显色；颜色变化后加入50μl 2M硫酸终止反应，之后于酶标仪OD450nm-620nm通过酶标仪进行分析。效价检测结果见表1。
表1各BALB/c小鼠免疫效价
(3)杂交瘤细胞的融合
根据表1的检测结果，选取具有抗重组人IL-1β免疫球蛋白较高滴度的小鼠进行融合，融合前先对小鼠作抗原腹腔加强免疫，连续加强免疫3天后，处死小鼠并取出脾脏，然后利用电融合法将分离自BALB/c小鼠的小鼠脾细胞与小鼠骨髓瘤细胞系融合(共3次融合)，获得能够分泌表达抗人IL-1β鼠多抗的杂交瘤细胞。融合方法包括：
将来自小鼠脾脏的B细胞与sp2/0细胞按1:1-1:2的比例混匀，离心之后去上清，用10ml Medium C(BTX,47-0001)重悬细胞清洗2次，之后用Medium C调整细胞密度为1×107/ml进行电融合。
将融合后的细胞以2×104/孔铺在96孔板，之后在选择性培养基中温育4-7天，该选择性培养基是在RPMI Medium 1640(Gibco,11875-093)加入终浓度为1×Penicillin-Streptomycin(Gibco,15140122),终浓度为1×GlutaMax-100×(Gibco,35050-061)，终浓度为1×HAT(Gibco,21060-017)，终浓度为1×HyMax-20×(Gibco,113004)，以及终浓度为20％的胎牛血清；4-7天后用HT替换HAT进行培养。
(4)亚克隆筛选
杂交瘤细胞培养10-14天后，通过ELISA(方法步骤参考步骤(2))对96孔板各个孔内的培养液进行IL-1β鼠多抗筛选；然后利用有限稀释法，将对应阳性孔内的母克隆(OD＞0.2)按照0.8/孔的细胞数目进行亚克隆铺板；10天后检测杂交瘤细胞培养基上清，将对应阳性孔的亚克隆(OD>0.2)转移至24孔板中生成小量抗体用于进一步表征。
(5)抗IL-1β杂交瘤上清的体外功能筛选
根据IL-1β可刺激MRC-5(人胚胎成纤维细胞)产生IL-6的原理，应用ELISA方法检测IL-6的表达水平来间接反映抗体的阻断功能。
当MRC-5细胞达到60％的汇合度时，胰酶消化，之后用培养基重悬调整MRC-5细胞密度为2×104；在96孔平底板中加入50μl的MRC-5细胞，之后加入25μl的杂交瘤细胞上清(即步骤(4)中筛选出的亚克隆)以及对照抗体(所有样本作1:1和1:10两个稀释倍数)，加入25μl稀释过的hIL-1β，于37℃、5％CO2培养箱中孵育18-24小时；之后收集上清，用ELISA试剂盒检测IL-6的分泌水平，在540nm处读取吸光度值。检测结果见图1和图2。
由图1和图2可见，各亚克隆杂交瘤细胞所分泌的抗体均表现出不同的阻断功能，本实施例选取其中阻断功能较好的A13-8、A13-60、B8-8、B27-4和C20-2亚克隆进行基因测序以及抗IL-1β人鼠嵌合抗体的构建。
实施例2抗IL-1β人鼠嵌合抗体的构建及表达纯化
收集扩大培养的亚克隆杂交瘤细胞，提取mRNA，逆转录成cDNA，经PCR扩增、克隆和测序，分别获得A13-8、A13-60、B8-8、B27-4和C20-2亚克隆表达的抗IL-1β鼠单抗的可变区序列如表2所示：
表2各抗IL-1β鼠单抗的可变区序列
在各可变区序列前添加信号肽序列(如SEQ ID No.48所示)，并在重链可变区序列后添加重链恒定区序列(如SEQ ID No.34所示)，在轻链可变区序列后添加轻链恒定区序列(如SEQ ID No.35所示)，然后在重链恒定区序列和轻链恒定区序列后添加TGA终止密码子；根据设计的重链和轻链氨基酸序列合成相应的基因序列，并将合成的基因序列构建至真核细胞表达载体中。
阳性对照抗体CAN的重轻链氨基酸序列来自美国专利US20040063913，具体的序列信息为SEQ ID No.36和SEQ ID No.37所示，同样地，在CAN氨基酸序列前添加信号肽、氨基酸序列后添加TGA终止密码子后进行对应的基因序列合成与真核细胞表达载体构建。基因合成与载体构建由苏州泓迅生物科技股份有限公司完成。
将构架的载体质粒组合(即包含抗体重链的载体质粒和包含抗体轻链的载体质粒)瞬时转染293细胞，培养7天后，利用亲和层析的方法纯化和富集细胞上清中的抗体。
载体质粒扩大后采用从天根生化科技(北京)有限公司购买的质粒提取试剂盒进行质粒抽提，获得的载体质粒用于如下的抗体组合表达。取轻链表达载体和重链表达载体以1:1比例混合后，加入稀释过的3倍体积(DNA加入的体积量)的PEI转染试剂。形成DNA-PEI复合物后滴加到293细胞中。转染24小时后，添加10％体积的生长培养基。7天后收取细胞上清，4000rpm离心20分钟后，取上清，再用0.22μm滤膜过滤，进行Protein A亲和层析纯化。纯化之后用PBS透析，4℃透析过夜。
实施例3抗IL-1β人鼠嵌合抗体亲和力的测定及种属交叉反应测定
本实施例利用Biacore分析验证抗IL-1β人鼠嵌合抗体的结合亲和力和结合动力学，以及其与猴IL-1β(Sino Biological Inc,90010-CNAE)的结合亲和力和结合动力学。
将嵌合抗体连接到Protein A芯片，通过将hIL-1β、猴IL-1β以5nm、50nm的浓度、30μl/min的流速在HBS-EP缓冲液中流动测量结合。使用BIAevaluation软件将结合和解离曲线拟合至1:1朗格谬尔(Langmuir)结合模型。测定的KD、Ka、Kd值显示于表3。
表3抗IL-1β人鼠嵌合抗体的Biacore结合数据
如上表所示，实施例2表达纯化的抗IL-1β人鼠嵌合抗体与人IL1β蛋白和猴IL1β蛋白均具有很高的亲和力。
实施例4抗IL-1β人鼠嵌合抗体的体外IC50测定
根据IL-1β可刺激MRC-5(人胚胎成纤维细胞)产生IL-6的原理，本实施例通过应用ELISA试剂盒检测IL-6的分泌水平来间接反映抗IL-1β人鼠嵌合抗体结合hIL-1β的能力。
当MRC-5细胞达到60％的汇合度时，胰酶消化，之后用培养基重悬调整其密度为2×104；在96孔平底板中加入50μl的MRC-5细胞，之后加入25μl 10倍梯度稀释的嵌合抗体以及对照抗体，加入25μl稀释过的hIL-1β；于37℃、5％CO2培养箱中孵育18-24小时，之后收集上清，用ELISA试剂盒检测IL-6的分泌水平，在540nm处读取吸光度值。检测结果如图3所示。
由图3可见，B27-4与C20-2表现出了和阳性对照抗体相当的功能抑制活性。
实施例5抗IL-1β抗体人源化改造
为了进一步提高B27-4抗体的空间构型稳定性，同时提高抗体的人源化程度，降低免疫副作用，本实施例对B27-4抗体作进一步改造。
关于抗体人源化改造有很多报道的成功案例，可供参考(Kettleborough C A,Saldanha J W,Heath V J,et al.Humanization of a mouse monoclonal antibody byCDR–grafting:the importance of framework residues on loop conformation[J].Protein Engineering,1991,4(7):773-783；Acqua W F,Damschroder M,Zhang J,etal.Antibody humanization by framework shuffling[J].Methods,2005,36(1):43-60)，本实施例采用抗体CDR移植和蛋白质同源建模的方法对B27-4抗体可变区进行抗体人源化改造，表达后使用Biacore分析改造后人源化抗体的亲和力，同时比较人源化后的抗体hB27-4与对照抗体CAN的亲和力。
经过同源建模和生物信息学分析后，人源化后抗体Hb27-4的信息如下表所示。
表4人源化抗IL-1β人鼠嵌合抗体的氨基酸序列信息
(注：SEQ ID No.1-48为氨基酸序列，其对应的核苷酸序列依次如SEQ IDNo.49-96所示。)
参考实施例2和实施例3的方法，对人源化抗体进行基因合成、载体构建和抗体表达，表达后的抗体上清进行Biacore仪器进行亲和力检测，使用BIAevaluation软件将结合和解离曲线拟合至1:1朗格谬尔(Langmuir)结合模型。测定的KD、Ka、Kd值显示于下表：
表5人源化抗体hB27-4与CAN抗体的Biacore结合数据
| 样品ID | 分析物 | ka(1/Ms) | kd(1/s) | KD(M) |
| hB27-4 | rHuman IL-1β | 8.29E+05 | 4.51E-04 | 5.45E-10 |
| CAN | rHuman IL-1β | 3.93E+06 | 6.01E-05 | 1.53E-11 |
由表5可见，改造后的抗体很好的保留了与人IL-1β蛋白的亲和力，并且亲和力水平与对照抗体相当。经过人源化改造的抗体hB27-4在保持现有的生物学功能情况下也会有更低的免疫原性、更低的副作用。
Claims (6)
1.一种人IL-1β蛋白结合分子，所述的人IL-1β蛋白结合分子具有与人IL-1β蛋白特异结合的抗原结合位点，该抗原结合位点含有至少一个重链可变区和至少一个轻链可变区，其特征在于，所述的轻链可变区的氨基酸序列如SEQ IDNo.44所示，所述的重链可变区的氨基酸序列如SEQ ID No.45所示。
2.如权利要求1所述的人IL-1β蛋白结合分子，其特征在于，为人源化的嵌合抗体、抗原结合片段或单链抗体可变区片段。
3.编码如权利要求1所述的人IL-1β蛋白结合分子的核苷酸分子。
4.含有如权利要求3所述的编码人IL-1β蛋白结合分子的核苷酸分子的表达载体。
5.如权利要求1-2中任意一项所述的人IL-1β蛋白结合分子在制备IL-1介导型疾病治疗药物中的应用，其特征在于，所述的疾病为自身免疫病或癌症。
6.如权利要求1-2中任意一项所述的人IL-1β蛋白结合分子在制备双特异性抗体中的应用，其特征在于，所述的人IL-1β蛋白结合分子为人源化的抗原结合片段或单链抗体可变区片段。
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