CN111487207A - 6-磷酸葡萄糖脱氢酶突变体及其在制备地高辛检测试剂中的用途 - Google Patents
6-磷酸葡萄糖脱氢酶突变体及其在制备地高辛检测试剂中的用途 Download PDFInfo
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- CN111487207A CN111487207A CN202010004879.2A CN202010004879A CN111487207A CN 111487207 A CN111487207 A CN 111487207A CN 202010004879 A CN202010004879 A CN 202010004879A CN 111487207 A CN111487207 A CN 111487207A
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Abstract
本申请涉及6‑磷酸葡萄糖脱氢酶突变体及其在制备地高辛检测试剂中的用途。具体而言,本申请的6‑磷酸葡萄糖脱氢酶突变体相较于野生型6‑磷酸葡萄糖脱氢酶包含选自以下的一个突变或其组合:D306C、D375C、G426C。使用本申请的6‑磷酸葡萄糖脱氢酶突变体所制备的检测试剂盒,其特异性强、灵敏度高、操作方便、检测时间短、定量准确,适合高通量检测。
Description
本申请要求2019年1月9日提交的201910017764.4和2019年5月21日提交的201910423122.4《6-磷酸葡萄糖脱氢酶突变体及其在制备检测试剂中的用途》的优先权,其通过引用并入此处。
技术领域
本申请涉及生物检测领域,特别是涉及一种突变的酶6-磷酸葡萄糖脱氢酶(简称G6PDH)及其在地高辛检测试剂盒中的应用。
背景技术
半抗原,某些小分子物质(分子量小于4000Da),其单独不能诱导免疫应答,即不具备免疫原性,但当其与大分子蛋白质或非抗原性的多聚赖氨酸等载体交联或结合后可获得免疫原性,诱导免疫应答。这些小分子物质可与应答效应产物结合,具备抗原性,它只有免疫反应性,不具免疫原性,又称不完全抗原。
半抗原能与对应抗体结合出现抗原-抗体反应,又不能单独激发人或动物体产生抗体的抗原。它只有免疫反应性,不具免疫原性,又称不完全抗原。大多数多糖、类脂、激素、小分子药物都属于半抗原。如果用化学方法把半抗原与某种蛋白分子(载体)结合,会获得新的免疫原性,并能刺激动物产生相应的抗体。半抗原一旦与蛋白结合,就构成该蛋白质的一个抗原簇。一些比一般半抗原分子量小,但有特异结构的化学活性基团物质(如青霉素、磺胺剂等),称为简单半抗原。
小分子抗原或半抗原因缺乏可作夹心法的两个以上的位点,因此不能用双抗体夹心法进行测定,多采用竞争模式。原理是标本中的抗原和一定量的酶标抗原竞争与固相抗体结合。标本中抗原量含量愈多,结合在固相上的酶标抗原愈少,显色愈浅。小分子激素、药物等ELISA测定多用此法。
地高辛(Digoxin)结构式如下所示:
地高辛是强心苷药物,对心脏有正性肌力作用,减慢心率,抑制心脏传导,用于治疗高血压、瓣膜性心脏病、先天性心脏病等急性和慢性心功能不全。尤其适用于伴有快速心室率的心房颤动的心功能不全。
常见的不良反应包括:促心律失常作用、恶心、呕吐、下腹痛、异常的无力、软弱;少见的反应包括:视力模糊,中枢神经系统反应(如精神抑郁或错乱);罕见的反应包括:嗜睡、头痛及皮疹、寻麻疹(过敏反应)。在洋地黄的中毒表现中,促心律失常最重要,其次为房室传导阻滞、阵发性或加速性交界性心动过速、阵发性房性心动过速伴房室传导阻滞、室性心动过速、窦性停搏、心室颤动等。
因此,应对该药不良反应监测予以重视。而且由于药物代谢个体存在差异,临床使用时应结合血药浓度监测,制定合理的给药方案,尽量避免不良反应发生。
目前已知的地高辛检测方法主要有:高效液相色谱法(HPLC)、化学发光免疫、酶联免疫吸附剂测定(ELISA)、均相酶免疫测定法、胶乳凝集比浊法等方法。HPLC法需要复杂的样品前处理,操作复杂周期长且成本昂贵;发光免疫法试剂成本昂贵,不适合常规治疗药物检测,更不利于大范围推广。现有的均相酶免疫测定法、胶乳凝集比浊法常常因制备工艺复杂、批间差大,应用受到一定限制。
现有技术CN108593905A描述了一种地高辛检测试剂盒及其制备方法。然而,现有技术的方法依赖于对小分子药物(地高辛)自身所带反应基团进行的激活,之后再与酶进行反应。这样的偶联方法会出现同一葡萄糖六磷酸脱氢酶上链接有多个地高辛的情况,且偶联位点难以确保一致性,难以保证小分子药物和酶之间的定向1:1反应,而导致批间差异大。
发明内容
鉴于本领域的需求,本申请提供了一种新型的6-磷酸葡萄糖脱氢酶突变体、及其在制备地高辛检测试剂盒中的用途。
根据一些实施方案,提供了一种6-磷酸葡萄糖脱氢酶突变体。区别于已有发表的专利US006090567A(Homogeneous immunoassays using mutant glucose-6-phosphatedehydrogenases)的6磷酸葡萄糖脱氢酶的突变体,本申请的6-磷酸葡萄糖脱氢酶突变体,其包含选自以下的突变:D306C、D375C、G426C。
根据一些实施方案,提供了一种6-磷酸葡萄糖脱氢酶突变体,所述6-磷酸葡萄糖脱氢酶突变体是选自以下的序列所示:SEQ ID No.2、SEQ ID No.3、SEQ ID No.4。
根据一些实施方案,提供了一种多核苷酸,其编码本申请的6-磷酸葡萄糖脱氢酶突变体。
根据一些实施方案,提供了一种表达载体,其包含本申请的多核苷酸。
根据一些实施方案,提供了一种宿主细胞,其包含本申请的表达载体。宿主细胞可以是原核(如细菌)或真核(如酵母)。
根据一些实施方案,提供了一种偶联物,其是本申请的6-磷酸葡萄糖脱氢酶突变体与半抗原按照摩尔比1:n偶联而成。
在一些实施方案中,n是1至50,例如,1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、41、42、43、44、45、46、47、48、49、50。
在一些具体的实施方案中,本申请的6-磷酸葡萄糖脱氢酶突变体与半抗原按照摩尔比优选为1:1。
在一些具体的实施方案中,半抗原的分子量为100Da至4000Da,例如:100、150、200、250、300、350、400、410、420、430、440、450、460、470、480、490、500、520、550、570、600、620、650、700、750、800、850、900、950、1000、1100、1200、1300、1400、1500、1600、1700、1800、1900、2000、2100、2200、2300、2400、2500、2600、2700、2800、2900、3000、3100、3200、3300、3400、3500、3600、3700、3800、3900、4000。
根据本申请,技术人员将理解,“半抗原”还包含其衍生物的形式。为了便于和6-磷酸葡萄糖脱氢酶进行偶联,对于那些自身不带有偶联基团(例如,与巯基反应的基团)的半抗原(例如地高辛),可以经改造而带有接头,以便和巯基共价结合。因此,在本申请中,半抗原衍生物是指,经改造而带有巯基反应基团的半抗原。
半抗原选自:小分子药物(如抗生素、精神药物)、激素、代谢物、糖、脂质、氨基酸。
半抗原例如但不限于:茶碱、苯妥英、维生素D、25羟维生素D、1,25双羟维生素D、叶酸、强心甙(包括地高辛、洋地黄毒苷)、酶酚酸、雷帕明、环胞菌素A、乙胺碘复酮、甲胺喋呤、他克莫司、血清氨基酸、胆汁酸、甘胆酸、苯丙氨酸、乙醇、尿尼柯丁代谢产物柯替宁、尿吗啡、尿单羟酚衍生物、神经肽酪氨酸、血浆甘丙素、多胺、组织胺、促甲状腺激素、泌乳素、胎盘泌乳素、生长激素、促卵泡刺激素、促黄体生成素、促肾上腺皮质激素、抗利尿激素、降钙素、降钙素原、甲状旁腺激素、甲状腺素、三碘甲状原氨酸、反三碘甲状原氨酸、游离甲状腺素、游离三碘甲状原氨酸、皮质醇、尿17-羟皮质类固醇、尿17-酮类固醇、脱氢表雄酮及硫酸酯、醛固酮、尿香草苦杏仁酸、血浆肾素、血管紧张素、促红细胞生成素、睾酮、双氢睾酮、雄烯二酮、17α羟孕酮、雌酮、雌三醇、雌二醇、孕酮、人绒毛膜促性腺激素、胰岛素、胰岛素原、C肽、胃泌素、血浆前列腺素、血浆6-酮前列腺素F1α、前列环素、肾上腺素、儿茶酚胺、去甲肾上腺素、胆囊收缩素、纳素、环磷酸腺苷、环磷酸鸟苷、血管活性肽、生长抑素、促胰液素、P-物质、神经降压素、血栓素A2、血栓素B2、5羟色胺、神经肽Y、骨钙素。
在具体的实施方案中,半抗原是地高辛或其衍生物。
在具体的实施方案中,半抗原是地高辛衍生物,其带有巯基反应基团,例如来酰亚胺、溴乙酰基、乙烯基砜或氮丙啶。
在具体的实施方案中,半抗原是地高辛衍生物,如式I所示:
其中,
在一些实施方案中,m为1至10的整数,优选1至5的整数,例如1、2、3、4、5。
在一些具体的实施方案中,地高辛衍生物具有式I-1所示的结构:
其中,
根据一些实施方案,提供了一种试剂,其包含本申请的偶联物。
根据一些实施方案,提供了本申请的6-磷酸葡萄糖脱氢酶突变体在制备地高辛检测试剂中的用途。
根据一些实施方案,提供了本申请的偶联物在制备地高辛检测试剂中的用途。
在具体的实施方案中,所述的检测试剂选自:酶联免疫法检测试剂、化学发光免疫法检测试剂、均相酶免疫法检测试剂、胶乳增强免疫比浊法检测试剂。
在具体的实施方案中,所述的检测试剂优选是基于竞争法检测的试剂。
根据一些实施方案,提供了本申请的偶联物在制备地高辛检测装置中的用途。
在具体的实施方案中,所述的检测装置可以制备成孔板(例如96-孔板)的形式,比如板上包被根据本申请的试剂。
在具体的实施方案中,所述的检测装置可以制备成颗粒(例如胶乳、磁珠)的形式,比如颗粒上包被根据本申请的试剂。
根据一些实施方案,提供了一种地高辛检测试剂盒,其包含:
-第一试剂,所述第一试剂包含底物、缓冲液和地高辛抗体;所述底物是6-磷酸葡萄糖脱氢酶的底物;
-第二试剂,所述第二试剂包含本申请的偶联物和缓冲液;
-任选地,校准品,所述校准品包含10mM至500mM缓冲液、0ng/ml至5ng/ml地高辛(例如0、0.5、1、1.5、2、2.5、3、3.5、4、4.5、5ng/ml或之间的任意值);以及
-任选地,质控品,所述质控品包含10mM至500mM缓冲液、0.7ng/ml至4.5ng/ml(例如0.7、0.8、1.5、1.8、2、3、4、4.5ng/ml或之间的任意值)地高辛。
根据一个实施方案,提供了一种地高辛检测试剂盒,其包含:
第一试剂,其包含:
10mM至500mM缓冲液、
5mM至50mM底物、
0.01μg/ml至10μg/ml地高辛抗体(0.1、0.2、0.3、0.4、0.5、0.6、0.7、0.8、0.9、1.0、1.1、1.2、1.5、2、3、4、5μg/ml)、
0.1g/L至5g/L稳定剂、
0.1g/L至5g/L表面活性剂、
0.1g/L至5g/L防腐剂;
第二试剂,其包含:
10mM至500mM缓冲液、
0.01μg/ml至10μg/ml根据本申请的偶联物(0.05、0.06、0.07、0.08、0.09、0.1、0.2、0.3、0.4、0.5、0.6、0.7、0.8、0.9、1.0μg/ml)、
0.1g/L至5g/L稳定剂、
0.1g/L至5g/L表面活性剂、
0.1g/L至5g/L防腐剂。
在一些实施方案中,所述缓冲液选自以下的一种或组合:TAPS、氨基丁三醇缓冲液、磷酸盐缓冲液、Tris-HCl缓冲液、柠檬酸-柠檬酸钠缓冲液、巴比妥缓冲液、甘氨酸缓冲液、硼酸盐缓冲液、三羟甲基甲烷缓冲液;优选,磷酸盐缓冲液;所述缓冲液的浓度为10mmol/L至500mmol/L,优选50至100mM;所述缓冲液的pH为7至8。
在一些实施方案中,所述稳定剂选自以下的一种或组合:牛血清白蛋白、海藻糖、甘油、蔗糖、甘露醇、甘氨酸、精氨酸、聚乙二醇6000、聚乙二醇8000;优选牛血清白蛋白。
在一些实施方案中,所述表面活性剂选自以下的一种或组合:Brij23、Brij35、Triton X-100、Triton X-405、Tween20、Tween30、Tween80、椰子油脂肪酸二乙醇酰胺、AEO7,优选Tween20。
在一些实施方案中,所述的防腐剂选自以下的一种或组合:叠氮化物、MIT、生物防腐剂PC(如PC-300)、硫柳汞;所述叠氮化物选自:叠氮钠、叠氮锂、PC-300。
在一些实施方案中,所述底物包含:6-磷酸葡糖糖、β-烟酰胺腺嘌呤二核苷酸。
在一些具体的实施方案中,所述地高辛抗体源自:小鼠、大鼠、猫、犬、灵长类、牛、马、羊、骆驼科、禽、人。
在一些具体的实施方案中,所述地高辛抗体选自:单抗、多抗、重组抗体、嵌合抗体、抗原结合片段。
根据一些实施方案,提供了一种偶联物的制备方法,包括步骤:
1)提供根据本申请的地高辛衍生物,尤其是在非质子性溶剂(例如但不限于乙腈、二甲基甲酰胺、二甲基亚砜)中提供根据本申请的地高辛衍生物;
2)提供6-磷酸葡萄糖脱氢酶突变体,优选在缓冲液(其提供反应环境,例如但不限于PBS、Tris、TAPS、TAPSO,所述缓冲液pH为6.0至8.0)中提供6-磷酸葡萄糖脱氢酶突变体;
3)在18℃至28℃,将所述6-磷酸葡萄糖脱氢酶突变体和所述地高辛衍生物,按照地高辛衍生物:酶=500:1至1:500的摩尔比(优选50:1至1:50)接触1小时至4小时(1、1.5、2、2.5、3、3.5、4小时、或之间的任何值,优选2小时至3小时)使得所述地高辛衍生物和所述6-磷酸葡萄糖脱氢酶突变体发生偶联,得到所述种偶联物;
4)根据需要,任选对所述种偶联物进行纯化,例如脱盐处理等。
在一些实施方案中,反应体系中半抗原和酶的接触摩尔比1:n,其中n是1至500,例如,1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、41、42、43、44、45、46、47、48、49、50、100、200、300、400、500及其任意上述数值之间的范围。
在一些具体的实施方案中,步骤1)和2)可互换或并行。
在一些具体的实施方案中,在偶联之前,所述6-磷酸葡萄糖脱氢酶包含一个或多个游离的巯基,从而允许和地高辛实现定向反应。
野生型6-磷酸葡萄糖脱氢酶不含游离的巯基,因此在一些具体的实施方案中,6-磷酸葡萄糖脱氢酶是经过基因工程改造的,使其在特定位点(306、375或426位)上的氨基酸突变为半胱氨酸,从而带有一个游离巯基。
附图说明
图1.G6PDH(野生型)氨基酸序列(SEQ ID No.1);源自明串珠菌属假肠膜明串珠菌Leuconostoc pseudomesenteroides。
图2.G6PDH(D306C)氨基酸序列(SEQ ID No.2)。
图3.G6PDH(D375C)氨基酸序列(SEQ ID No.3)。
图4.G6PDH(G426C)氨基酸序列(SEQ ID No.4)。
具体实施方式
实施例
实施例1.地高辛衍生物的合成
1.化合物2的合成
将1.0g地高辛溶于95%的乙醇(80ml)中,然后向其中加入高碘酸(1.0g)水(10ml)溶液,室温(18-28℃)搅拌1小时。过滤除去残渣,减压除去溶剂,二氯甲烷萃取。有机相使用无水硫酸钠干燥,减压除去溶剂得到化合物2(白色固体,0.99g,99%)。
2.化合物4的合成
将化合物2(900mg,1.16mmol)溶于10ml干燥的甲醇中,将化合物3(318mg,1.0mmol)加入反应体系,室温搅拌5分钟。加入氰基硼氢化钠(146mg,2.32mmol),室温搅拌约12小时。减压除去溶剂,直接柱层析纯化得到化合物4(白色固体,585mg,66%)。
3.化合物5的合成
将化合物4溶于15ml二氯甲烷中,氮气条件下室温搅拌30分钟,然后加入10ml哌啶,室温搅拌2小时。减压除去溶液,柱层析纯化得到化合物5(460mg,78%)。
4.地高辛衍生物的合成
化合物5(88mg,0.11mmol)和化合物6(17mg,0.11mmol)溶解于DCM(8mL)中,向其中滴加三乙胺(33mg,0.33mmol),然后加入HATU(50mg,0.13mmol),室温下搅拌2h,得到地高辛衍生物(白色固体,50mg,45%)。
5.经质谱和核磁鉴定,地高辛衍生物结构正确无误。
本实施例使得地高辛带有一个可以和酶结合的基团。
实施例2.地高辛衍生物与G6PDH分子的偶联
一、本申请的偶联方法
根据本申请的G6PDH-地高辛偶联物,按照以下方式进行偶联:地高辛衍生物分子上的巯基反应性基团(如但不限于马来酰亚胺基团)与G6PDH分子上的巯基共价结合。
1.将实施例1制备的地高辛衍生物溶于N,N-二甲基甲酰胺中(10mg/ml);
2.G6PDH溶液:G6PDH(例如本申请的突变体)溶于PB 100mmol、NaCl 100mmol、pH=8.0;
3.将200μl G6PDH溶液加入至750μl缓冲溶液(0.05M Na2HPO4、150mM NaCl、10mMEDTA、0.1%NaN3、pH=7.2)中;然后向其中加入地高辛衍生物的N,N-二甲基甲酰胺溶液50μl;
4.上述混合溶液,在室温(18-28℃)下充分震荡2-3小时,脱盐处理,收集蛋白峰,所得产物即G6PDH-地高辛偶联物。
二、对照偶联方法(参照CN108593905A的方法)
准确称取100-300mg地高辛,并用5-15mL无水乙醇溶解;
在上述溶液中滴加10-200mM的高碘酸钠5-15mL,并轻微振荡,室温搅拌反应0.5-2小时;
滴加0.5-2M的乙二醇0.5-1mL,室温搅拌反应5-10分钟;
将上述反应混合物滴加到5-15mL正在搅拌中的2-3%的G6PDH溶液中,并调节溶液pH到9.0-9.5,继续搅拌反应0.5-2小时,并使溶液pH稳定;
加入100-200mg四氢硼钠,搅拌还原12-24小时;
通过G-25凝胶层析柱纯化得到G6PDH-地高辛偶联物。
实施例3.试剂盒的制备
制备以下检测地高辛的试剂盒,其包含:
试剂R1,包含:
TAPS缓冲液100mM,pH 7.0
15mM 6-磷酸葡萄糖
15mMβ-烟酰胺腺嘌呤二核苷酸
0.5μg/ml地高辛抗体(市售抗体,无特殊限制)
1g/L牛血清白蛋白
1g/L Brij
1g/L叠氮钠;
试剂R2,包括:
磷酸缓冲液200mM,pH8.0
0.1μg/ml G6PDH-地高辛偶联物
100mM NaCl
1g/L牛血清白蛋白
1g/L Brij
1g/L叠氮钠;
校准品:20mM HEPES缓冲液,以及0ng/ml、0.5ng/ml、1ng/ml、2ng/ml、3ng/ml、5ng/ml地高辛(或按需加入);
质控品:20mM HEPES缓冲液,以及0.6-0.8ng/ml、1.6-2.0ng/ml、3.8-4.2ng/ml地高辛(或按需加入)。
将上述试剂(任选包含质控品、校准品),组装成地高辛均相酶免疫检测试剂盒。
检测例
均相酶免疫测定的原理:在液体均相反应体系中,酶标记抗原(如G6PDH-地高辛)与非标记抗原(地高辛),竞争与定量的抗体(地高辛抗体)进行结合,当抗体与非标记抗原结合越多,酶标记抗原释放的活性就越多,酶催化底物NAD+生成NADH就越多。
在340nm波长下检测NADH的吸光度变化,即可推算出液体中地高辛的含量。
表1.全自动生化仪参数
检测例1.本申请试剂盒的性能
1.定标实验
表2.地高辛检测试剂盒定标吸光度
2.精密度实验
表3.总不精密度
3.重复性
表4.重复性
4.回收测试
表5.回收数据
5.线性实验
表6.线性
检测例2.机载稳定性
本申请试剂(D375C突变体)37℃加速7天后,定标吸光度下降小于10%,对照试剂37℃加速7天后,定标吸光度下降显著。
表7.37℃加速稳定性
检测例3.抗体抑制率
1.抗体抑制率的检测原理
当抗体与G6PDH-地高辛偶联物结合时,由于空间位阻导致G6PDH酶活性受到影响,从而使得其催化NAD转化为NADH的效率降低,通过检测NADH量的变化,从而比较加入抗体与未加入抗体的实验组的差异,这种差异即体现为抗体对G6PDH的抑制能力。
2.反应体系
表8.抗体抑制率的检测试剂制备
3.结果
比较加入抗体与未加入抗体时,分别检测G6PDH-地高辛偶联物吸光度测值,即可得到抗体对G6PDH的抑制情况。
抗体抑制率=含抗体时G6PDH-地高辛偶联物的吸光度变化值/不含抗体时G6PDH-地高辛的吸光度变化值×100%。
相对于已发表的突变位点(A45C),本申请的突变体在酶活性保留方面有明显提高,能达到40%以上(G426C:40%;D375C:49%),最高达62%(D306C)。将已发表的突变位点(例如A45C、K55C)参照本申请的方法制备成G6PDH-地高辛偶联物,其抑制率仅为33%和40%。
虽然不限于具体理论,但是可以部分地解释为:和现有技术中的G6PDH突变体(A45C、K55C)相比,本申请酶突变体中突变位点(即引入游离巯基的位点)是和半抗原(比如激素、小分子药物等)发生偶联的位置所在。半抗原在这个位置上与半抗原特异性抗体结合时,所构成的空间位阻对G6PDH酶的活性影响最小,同时在引入突变后,还不能实质上影响分子的空间折叠。因此,这个突变位点的位置非常重要,需要同时兼顾G6PDH酶的活性、偶联分子的空间折叠、以及半抗原表位的充分暴露。
由于酶的突变体在抗体抑制率上有明显的提高。将酶的突变体与地高辛偶联后的偶联物配制成试剂盒后,试剂在批间变异系数、线性、重复性、稳定性等性能方面有明显的性能提升。
序列表
<110> 北京九强生物技术股份有限公司
<120> 6-磷酸葡萄糖脱氢酶突变体及其在制备地高辛检测试剂中的用途
<130> 390266CG
<150> 201910017764.4
<151> 2019-01-09
<150> 201910423122.4
<151> 2019-05-21
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Claims (9)
2.根据权利要求1所述的偶联物,其中:
所述6-磷酸葡萄糖脱氢酶突变体,相较于野生型6-磷酸葡萄糖脱氢酶,包含选自以下的一个突变:D306C、D375C、G426C;
优选地,所述6-磷酸葡萄糖脱氢酶突变体是选自以下的序列所示:SEQ ID No.2、SEQID No.3、SEQ ID No.4。
3.一种试剂,其包含权利要求1或2所述的偶联物。
4.权利要求1或2所述的偶联物在制备检测试剂中的用途,其中:
所述检测试剂是地高辛的检测试剂;
优选地,所述检测试剂选自:酶联免疫法检测试剂、化学发光免疫法检测试剂、均相酶免疫法检测试剂、胶乳增强免疫比浊法检测试剂。
5.权利要求1或2所述的偶联物在制备检测装置中的用途:
所述检测装置是针对地高辛的检测装置;
所述检测装置选自以下任一的形式:孔板、颗粒、芯片、试纸;
优选地,所述检测装置用于地高辛的均相免疫检测。
6.一种地高辛检测试剂盒,其包含:
第一试剂,所述第一试剂包含底物、地高辛抗体、缓冲液;
第二试剂,所述第二试剂包含权利要求1或2所述的偶联物、缓冲液;
任选地,校准品,所述校准品包含10mM至500mM缓冲液、0ng/ml至5ng/ml地高辛;以及
任选地,质控品,所述质控品包含10mM至500mM缓冲液、0.7ng/ml至4.5ng/ml地高辛。
7.根据权利要求6所述的地高辛检测试剂盒,其包含:
第一试剂,其包含:
10mM至500mM,优选50mM至300mM缓冲液、
5mM至50mM,优选10mM至20mM葡萄糖-6-磷酸、
5mM至50mM,优选10mM至20mM氧化型β-烟酰胺腺嘌呤二核苷酸、
0.01μg/ml至10μg/ml,优选0.1μg/ml至1μg/ml地高辛抗体、
0.1g/L至5g/L,优选1g/L至5g/L稳定剂、
0.1g/L至5g/L,优选1g/L至5g/L表面活性剂、
0.1g/L至5g/L,优选1g/L至5g/L防腐剂;
第二试剂,其包含:
10mM至500mM,优选50mM至300mM缓冲液、
0.01μg/ml至10μg/ml,优选0.05μg/ml至0.5μg/ml权利要求1或2所述的偶联物、
0.1g/L至5g/L,优选1g/L至5g/L稳定剂、
0.1g/L至5g/L,优选1g/L至5g/L表面活性剂、
0.1g/L至5g/L,优选1g/L至5g/L防腐剂;
所述缓冲液选自以下的一种或组合:TAPS缓冲液、磷酸盐缓冲液、甘氨酸缓冲液、Tris缓冲液、硼酸缓冲液、MOPS缓冲液、HEPES缓冲液;
所述缓冲液的pH为7至8;
所述稳定剂选自以下的一种或组合:牛血清白蛋白、海藻糖、甘油、蔗糖、甘露醇、甘氨酸、精氨酸、聚乙二醇6000、聚乙二醇8000;优选牛血清白蛋白;
所述表面活性剂选自以下的一种或组合:Brij23、Brij35、Triton X-100、Triton X-405、Tween20、Tween30、Tween80、椰子油脂肪酸二乙醇酰胺、AEO7,优选Tween20;
所述防腐剂选自以下的一种或组合:叠氮化物、MIT、生物防腐剂PC、硫柳汞;
优选,所述防腐剂选自:叠氮钠、叠氮锂、PC-300。
8.根据权利要求6所述的地高辛检测试剂盒,其包含:
第一试剂,其包含:
50mM TAPS缓冲液,pH 8.0、
15mM葡萄糖-6-磷酸、
15mM氧化型β-烟酰胺腺嘌呤二核苷酸、
0.5μg/ml地高辛抗体、
1g/L牛血清白蛋白、
1g/L Tween20、
1g/L叠氮钠;
第二试剂,其包含:
50mM Tris缓冲液,pH 8.0、
0.1μg/ml权利要求1或2所述的偶联物、
1g/L牛血清白蛋白、
1g/L Tween20、
1g/L叠氮钠。
9.一种偶联物的制备方法,其包括步骤:
1)提供地高辛或其衍生物;
2)提供权利要求2所限定的6-磷酸葡萄糖脱氢酶突变体;
3)所述6-磷酸葡萄糖脱氢酶突变体与所述地高辛或其衍生物发生偶联;
所述地高辛衍生物是式I所示:
其中,
m为1至10的整数,优选1至5的整数;
优选地,所述偶联物的制备方法,其包括步骤:
1)提供地高辛衍生物,优选在非质子性溶剂中提供地高辛衍生物;
2)提供权利要求2所限定的6-磷酸葡萄糖脱氢酶突变体,优选在缓冲液中提供所述6-磷酸葡萄糖脱氢酶突变体;
3)在18℃至28℃,将所述6-磷酸葡萄糖脱氢酶突变体和所述地高辛衍生物接触1小时至4小时,优选2小时至3小时,使得所述地高辛衍生物和所述6-磷酸葡萄糖脱氢酶突变体发生偶联,得到所述偶联物;
4)任选,对所述偶联物进行纯化,优选脱盐;
步骤1)和步骤2)能够互换或并行;
所述缓冲液选自:PBS、Tris、TAPS、TAPSO,
所述缓冲液pH为6.0至8.0;
所述非质子性溶剂选自以下一种或组合:乙腈、二甲基甲酰胺、二甲基亚砜;
优选,在步骤3)之前,所述6-磷酸葡萄糖脱氢酶突变体包含一个游离巯基;更优选,所述6-磷酸葡萄糖脱氢酶突变体在第306、375或426位有游离巯基。
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CN202310257027.8A Pending CN116144619A (zh) | 2019-01-09 | 2019-12-26 | 茶碱检测试剂盒 |
CN202211151405.6A Pending CN115791649A (zh) | 2019-01-09 | 2019-12-27 | 甘胆酸检测试剂盒 |
CN202211151264.8A Pending CN116008201A (zh) | 2019-01-09 | 2019-12-27 | 6-磷酸葡萄糖脱氢酶突变体及其在制备检测试剂中的用途 |
CN201911372535.0A Active CN112285038B (zh) | 2019-01-09 | 2019-12-27 | 6-磷酸葡萄糖脱氢酶突变体及其在制备洋地黄毒苷检测试剂中的用途 |
CN202310811210.8A Pending CN116626281A (zh) | 2019-01-09 | 2019-12-27 | 一种洋地黄毒苷检测试剂盒 |
CN201911372147.2A Active CN112285037B (zh) | 2019-01-09 | 2019-12-27 | 6-磷酸葡萄糖脱氢酶突变体及其在制备检测试剂中的用途 |
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CN202310811212.7A Pending CN116735512A (zh) | 2019-01-09 | 2019-12-27 | 偶联物在制备检测试剂中的用途 |
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CN202310364266.3A Pending CN116718764A (zh) | 2019-01-09 | 2019-12-31 | 偶联物在制备苯妥英检测试剂中的用途 |
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CN202310726498.9A Pending CN116559472A (zh) | 2019-01-09 | 2019-12-31 | 一种皮质醇检测试剂盒 |
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CN202310217235.5A Pending CN116359146A (zh) | 2019-01-09 | 2019-12-26 | 偶联物的制备方法 |
CN201911365439.3A Active CN111239060B (zh) | 2019-01-09 | 2019-12-26 | 6-磷酸葡萄糖脱氢酶突变体及其在制备茶碱检测试剂中的用途 |
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CN201911372535.0A Active CN112285038B (zh) | 2019-01-09 | 2019-12-27 | 6-磷酸葡萄糖脱氢酶突变体及其在制备洋地黄毒苷检测试剂中的用途 |
CN202310811210.8A Pending CN116626281A (zh) | 2019-01-09 | 2019-12-27 | 一种洋地黄毒苷检测试剂盒 |
CN201911372147.2A Active CN112285037B (zh) | 2019-01-09 | 2019-12-27 | 6-磷酸葡萄糖脱氢酶突变体及其在制备检测试剂中的用途 |
CN202310811498.9A Pending CN116698772A (zh) | 2019-01-09 | 2019-12-27 | 偶联物的制备方法 |
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CN202310726493.6A Pending CN116773795A (zh) | 2019-01-09 | 2019-12-31 | 一种偶联物的制备方法 |
CN202310726498.9A Pending CN116559472A (zh) | 2019-01-09 | 2019-12-31 | 一种皮质醇检测试剂盒 |
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CN201911404154.6A Active CN111504920B (zh) | 2019-01-09 | 2019-12-31 | 6-磷酸葡萄糖脱氢酶突变体及其在制备皮质醇检测试剂中的用途 |
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CN202310508217.2A Pending CN116298330A (zh) | 2019-01-09 | 2020-01-07 | 偶联物在制备检测试剂中的用途 |
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CN110174363A (zh) * | 2019-01-09 | 2019-08-27 | 北京九强生物技术股份有限公司 | 6-磷酸葡萄糖脱氢酶突变体及其在制备检测试剂中的用途 |
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JP2023509037A (ja) * | 2020-01-07 | 2023-03-06 | 北京九▲強▼生物技▲術▼股▲フン▼有限公司 | グルコース-6-リン酸デヒドロゲナーゼ変異体及び検出試薬の調製におけるその使用 |
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CN115236216B (zh) * | 2022-06-07 | 2024-03-01 | 合肥和合医疗科技有限公司 | 高效液相色谱串联质谱检测全血中免疫抑制剂的试剂盒,其制备方法和检测方法 |
Citations (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4190496A (en) * | 1971-05-14 | 1980-02-26 | Syva Company | Homogeneous enzyme assay for antibodies |
US4341866A (en) * | 1980-06-02 | 1982-07-27 | Syva Company | Antienzyme termination in enzyme immunoassays |
US4469797A (en) * | 1982-09-23 | 1984-09-04 | Miles Laboratories, Inc. | Digoxigenin immunogens, antibodies, labeled conjugates, and related derivatives |
US4622294A (en) * | 1985-02-08 | 1986-11-11 | Kung Viola T | Liposome immunoassay reagent and method |
WO1990014594A1 (en) * | 1989-05-23 | 1990-11-29 | Pharmacia Diagnostics Inc. | Homogeneous immunochemical method for determining haptens by means of ion selective electrodes |
US5068198A (en) * | 1986-03-26 | 1991-11-26 | Syntex (U.S.A.) Inc. | Liquid single reagent for assays involving confining gels |
JPH04144679A (ja) * | 1990-10-03 | 1992-05-19 | Sanyo Chem Ind Ltd | 酵素標識ハプテンの製法 |
US6033890A (en) * | 1993-04-08 | 2000-03-07 | Behring Diagnostics Gmbh | Homogeneous immunoassays using mutant glucose-6-phosphate dehydrogenases |
US20060046273A1 (en) * | 2004-08-27 | 2006-03-02 | Lin-Zhi International Inc. | Homogeneous enzyme immunoassay for oral fluid |
CN108593905A (zh) * | 2017-12-22 | 2018-09-28 | 太原瑞盛生物科技有限公司 | 一种地高辛免疫检测试剂及其制备和检测方法 |
CN110174363A (zh) * | 2019-01-09 | 2019-08-27 | 北京九强生物技术股份有限公司 | 6-磷酸葡萄糖脱氢酶突变体及其在制备检测试剂中的用途 |
Family Cites Families (51)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
FR2116301B1 (zh) * | 1970-12-07 | 1974-08-30 | Brun Lab Sa Le | |
US3997525A (en) * | 1974-01-16 | 1976-12-14 | Bio-Tec, Inc. | Tetra-125 iodo-di-tyramine of digitalis derivative and process for making the same |
DE2901218A1 (de) * | 1979-01-13 | 1980-07-17 | Byk Gulden Lomberg Chem Fab | Ung von theophyllin |
JPS5618983A (en) * | 1979-07-25 | 1981-02-23 | Eisai Co Ltd | Theophylline derivative and its preapration |
US4262089A (en) * | 1980-04-07 | 1981-04-14 | Syva Company | Theophylline antigens and antibodies |
JPS57178159A (en) * | 1981-04-27 | 1982-11-02 | Banyu Pharmaceut Co Ltd | Chemical reagent for detection of amicacin and its determination |
US4608336A (en) * | 1981-08-27 | 1986-08-26 | Miles Laboratories, Inc. | #3B theophylline immunoassay employing 9-theophylline reagents |
DE3483620D1 (de) * | 1983-03-11 | 1991-01-03 | Fujirebio Kk | Verfahren zur bestimmung von liganden. |
IT1199088B (it) * | 1984-03-09 | 1988-12-30 | Miles Italiana | Saggio di legame specifico mediante impiego di anti-g6pdh come marcante |
DE3919915A1 (de) * | 1989-06-19 | 1990-12-20 | Boehringer Mannheim Gmbh | Aminoalkylmaleimide und davon abgeleitete hapten- und antigenderivate sowie konjugate mit peptiden oder proteinen |
ATE193600T1 (de) * | 1990-11-20 | 2000-06-15 | Dade Behring Marburg Gmbh | Verfahren zur stabilisierung von enzym-konjugaten |
ES2091301T3 (es) * | 1990-11-20 | 1996-11-01 | Behringwerke Ag | Inmunoensayo de ciclosporina. |
CA2087397A1 (en) * | 1992-01-22 | 1993-07-23 | Kazuhisa Kubotsu | Immunoassay and reagents used therefor |
CA2156397C (en) * | 1993-04-08 | 2007-05-15 | Valerie Quesniaux | Rapamycin assay |
US5747352A (en) * | 1994-05-23 | 1998-05-05 | Beckman Instruments, Inc. | Reagents and methods for the rapid and quantitative assay of pharmacological agents |
EP1121383A1 (en) * | 1998-10-09 | 2001-08-08 | Isotechnika, Inc. | Methods for the production of antibodies to specific regions of cyclosporine and cyclosporine metabolites |
AUPP751398A0 (en) * | 1998-12-04 | 1999-01-07 | Commonwealth Scientific And Industrial Research Organisation | Methotrexate derivatives |
US7078495B1 (en) * | 1999-08-03 | 2006-07-18 | Dade Behring Inc. | Monoclonal antibodies to tacrolimus and immunoassay methods for tacrolimus |
US6653456B2 (en) * | 2001-07-31 | 2003-11-25 | Roche Diagnostics Corporation | Site-specific aminoglycoside derivatives and their use in immunodiagnostic assays |
US20050176080A1 (en) * | 2004-02-10 | 2005-08-11 | Vani Bodepudi | Hapten, immunogens and derivatives of ascomycin useful for preparation of antibodies and immunoassays |
TW200635954A (en) * | 2004-12-17 | 2006-10-16 | Isotechnika Inc | Metabolites of cyclosporin analogs |
US7189582B2 (en) * | 2005-04-27 | 2007-03-13 | Dade Behring Inc. | Compositions and methods for detection of sirolimus |
JP4746926B2 (ja) * | 2005-06-29 | 2011-08-10 | シスメックス株式会社 | グルコース−6−リン酸脱水素酵素含有試薬及びグルコース−6−リン酸脱水素酵素安定化方法 |
CN101638640B (zh) * | 2009-09-07 | 2011-01-12 | 北京利德曼生化股份有限公司 | 葡萄糖-6-磷酸脱氢酶及其核苷酸序列、重组载体、重组宿主细胞和试剂盒 |
CN103547679A (zh) * | 2010-11-24 | 2014-01-29 | Dh科技发展私人贸易有限公司 | 葡萄糖-6-磷酸脱氢酶的高产量、灵敏检测 |
CN102565399B (zh) * | 2010-12-07 | 2015-06-03 | 北京望尔生物技术有限公司 | 一种检测氢化可的松的方法及其专用酶联免疫试剂盒 |
CN102807618A (zh) * | 2011-08-10 | 2012-12-05 | 重庆金域医学检验所有限公司 | 苯妥因均相酶免疫检测试剂盒及其多克隆抗体的制备方法 |
JP5896375B2 (ja) * | 2011-09-09 | 2016-03-30 | 池田食研株式会社 | 改変型グルコース脱水素酵素遺伝子 |
CN102424829B (zh) * | 2011-10-26 | 2013-10-16 | 苏州汉酶生物技术有限公司 | 一种酶催化合成坦西莫司的方法 |
US8771964B2 (en) * | 2012-02-02 | 2014-07-08 | Siemens Healthcare Diagnostics Inc. | Compositions and methods for detection of methadone metabolite |
KR20150023729A (ko) * | 2012-06-14 | 2015-03-05 | 암브룩스, 인코포레이티드 | 핵 수용체 리간드 폴리펩티드에 접합된 항-psma 항체 |
CN103242446A (zh) * | 2012-07-25 | 2013-08-14 | 苏州博源医疗科技有限公司 | 茶碱免疫原及其制备方法和应用 |
CN102757391B (zh) * | 2012-08-01 | 2015-08-26 | 苏州博源医疗科技有限公司 | 一种苯巴比妥衍生物及其制备方法和应用 |
WO2015094852A2 (en) * | 2013-12-17 | 2015-06-25 | Siemens Healthcare Diagnostics Inc. | Preparation of multi-hapten mutant g6pdh conjugates and their use for detection of multiple analytes |
CN104016923B (zh) * | 2014-01-08 | 2016-08-31 | 南开大学 | 苯妥英衍生物及其制备方法和用途 |
CN103760348B (zh) * | 2014-02-11 | 2015-03-11 | 苏州博源医疗科技有限公司 | 一种甘胆酸免疫检测试剂及其制备和检测方法 |
CN103739703B (zh) * | 2014-02-11 | 2015-07-15 | 苏州博源医疗科技有限公司 | 甘胆酸免疫原、抗甘胆酸特异性抗体及检测试剂 |
JP6398295B2 (ja) * | 2014-04-30 | 2018-10-03 | ニプロ株式会社 | 変異型グルコース−6−リン酸脱水素酵素 |
CN104447745B (zh) * | 2014-11-06 | 2016-03-30 | 济南金域医学检验中心有限公司 | 一种茶碱均相酶免疫检测验试剂盒及其制备方法 |
CN104569373B (zh) * | 2015-01-27 | 2016-08-17 | 苏州博源医疗科技有限公司 | 一种甲氨蝶呤均相酶免疫检测试剂及其制备和检测方法 |
US10330683B2 (en) * | 2015-02-04 | 2019-06-25 | Genentech, Inc. | Mutant smoothened and methods of using the same |
CN105131105A (zh) * | 2015-07-27 | 2015-12-09 | 苏州博源医疗科技有限公司 | 皮质醇免疫原、衍生物、抗体、检测试剂及制备方法 |
CN106565809B (zh) * | 2016-07-08 | 2018-05-01 | 北京九强生物技术股份有限公司 | 一种β半乳糖苷酶的酶供体偶联物及其在甘胆酸检测中的用途 |
CN106226512B (zh) * | 2016-07-29 | 2018-10-16 | 胡清 | 一种试剂盒、试剂盒的制备方法及利用该试剂盒实现的外周血甘氨胆酸的检测方法 |
CN106190996B (zh) * | 2016-08-30 | 2019-05-21 | 美康生物科技股份有限公司 | 一种葡萄糖6磷酸脱氢酶突变体 |
CN109613251B (zh) * | 2016-09-22 | 2021-01-26 | 北京九强生物技术股份有限公司 | 大肠杆菌β半乳糖苷酶受体的用途 |
CN106872681B (zh) * | 2017-01-23 | 2019-11-19 | 四川精卫食品检测科技有限公司 | 丁胺卡那霉素和卡那霉素二合一快速检测酶联免疫试剂盒及其应用 |
CN108586562B (zh) * | 2018-05-08 | 2019-09-06 | 苏州博源医疗科技有限公司 | 一种皮质醇衍生物及其制备方法与应用 |
CN108717117A (zh) * | 2018-05-23 | 2018-10-30 | 太原瑞盛生物科技有限公司 | 一种万古霉素免疫检测试剂及其制备和检测方法 |
CN109111494A (zh) * | 2018-08-30 | 2019-01-01 | 苏州博源医疗科技有限公司 | 雌二醇衍生物、免疫原、抗体、酶标偶联物、检测试剂及其制备方法 |
CN112574969A (zh) * | 2020-12-28 | 2021-03-30 | 郑州伊美诺生物技术有限公司 | G6pdh突变体及其应用 |
-
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- 2019-05-21 CN CN201910423122.4A patent/CN110174363A/zh active Pending
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Patent Citations (12)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4190496A (en) * | 1971-05-14 | 1980-02-26 | Syva Company | Homogeneous enzyme assay for antibodies |
US4341866A (en) * | 1980-06-02 | 1982-07-27 | Syva Company | Antienzyme termination in enzyme immunoassays |
US4469797A (en) * | 1982-09-23 | 1984-09-04 | Miles Laboratories, Inc. | Digoxigenin immunogens, antibodies, labeled conjugates, and related derivatives |
US4622294A (en) * | 1985-02-08 | 1986-11-11 | Kung Viola T | Liposome immunoassay reagent and method |
US5068198A (en) * | 1986-03-26 | 1991-11-26 | Syntex (U.S.A.) Inc. | Liquid single reagent for assays involving confining gels |
WO1990014594A1 (en) * | 1989-05-23 | 1990-11-29 | Pharmacia Diagnostics Inc. | Homogeneous immunochemical method for determining haptens by means of ion selective electrodes |
JPH04144679A (ja) * | 1990-10-03 | 1992-05-19 | Sanyo Chem Ind Ltd | 酵素標識ハプテンの製法 |
US6033890A (en) * | 1993-04-08 | 2000-03-07 | Behring Diagnostics Gmbh | Homogeneous immunoassays using mutant glucose-6-phosphate dehydrogenases |
US20060046273A1 (en) * | 2004-08-27 | 2006-03-02 | Lin-Zhi International Inc. | Homogeneous enzyme immunoassay for oral fluid |
CN101048660A (zh) * | 2004-08-27 | 2007-10-03 | 灵芝国际股份有限公司 | 口腔液的同质酶免疫分析 |
CN108593905A (zh) * | 2017-12-22 | 2018-09-28 | 太原瑞盛生物科技有限公司 | 一种地高辛免疫检测试剂及其制备和检测方法 |
CN110174363A (zh) * | 2019-01-09 | 2019-08-27 | 北京九强生物技术股份有限公司 | 6-磷酸葡萄糖脱氢酶突变体及其在制备检测试剂中的用途 |
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