[发明专利]肿瘤治疗性疫苗CTP37CRM197免疫原及其制备方法与应用

摘要

本发明涉及一种免疫原及其制备方法，是以白喉毒素突变体CRM197为载体蛋白，通过异型双功能交联剂ε－马来酰亚胺基己酸N－羟基琥珀酰亚胺酯(EMCS)与人绒毛膜促性腺激素β亚基C末端37的氨基酸分子组成的肽(hCG－CTP37)相交联而得的CTP37CRM197。本发明所使用的载体蛋白具有回收率高，易于纯化，适合于大规模工业化生产的优点，同时交联产物交联率高，制备时间短，降低了生产成本。本发明也涉及用此方法制备的免疫原作为治疗性疫苗的有效成分，以适宜的免疫佐剂作为辅料制成的药物在肿瘤治疗中的应用。

主权项

1、一种免疫原，其特征是以白喉毒素突变体CRM197为载体蛋白，通过异型双功能交联剂ε-马来酰亚胺基己酸N-羟基琥珀酰亚胺酯(EMCS)与人绒毛膜促性腺激素β亚基C末端37个氨基酸分子组成的肽(hCG-CTP37)分子相交联而得的CTP37CRM197，其中，所述的CTP37分子的氨基酸序列如下：Thr-Cys-Asp-Asp-Pro-Arg-Phe-Gln-Asp-Ser-Ser-Ser-Ser-Lys-Ala-Pro-Pro-Pro-Ser-Leu-Pro-Ser-Pro-Ser-Arg-Leu-Pro-Gly-Pro-Ser-Asp-Thr-Pro-Ile-Leu-Pro-Gln所述的白喉毒素突变体CRM197的氨基酸序列如下：Met Gly Ala Asp Asp Val Val Asp Ser Ser Lys Ser Phe Val Met Glu Asn Phe Ser Ser1 5 10 15 20Tyr His Gly Thr Lys Pro Gly Tyr Val Asp Ser Ile Gln Lys Gly Ile Gln Lys Pro Lys21 25 30 35 40Ser Gly Thr Gln Gly Asn Tyr Asp Asp Asp Trp Lys Glu Phe Tyr Ser Thr Asp Asn Lys41 45 50 55 60Tyr Asp Ala Ala Gly Tyr Ser Val Asp Asn Glu Asn Pro Leu Ser Gly Lys Ala Gly Gly61 65 70 75 80Val Val Lys Val Thr Tyr Pro Gly Leu Thr Lys Val Leu Ala Leu Lys Val Asp Asn Ala81 85 90 95 100Glu Thr Ile Lys Lys Glu Leu Gly Leu Ser Leu Thr Glu Pro Leu Met Glu Gln Val Gly101 105 110 115 120Thr Glu Glu Phe Ile Lys Arg Phe Gly Asp Gly Ala Ser Arg Val Val Leu Ser Leu Pro121 125 130 135 140Phe Ala Glu Gly Ser Ser Ser Val Glu Tyr Ile Asn Asn Trp Glu Gln Ala Lys Ala Leu141 145 150 155 160Ser Val Glu Leu Glu Ile Asn Phe Glu Thr Arg Gly Lys Arg Gly Gln Asp Ala Met Tyr161 165 170 175 180Glu Tyr Met Ala Gln Ala Cys Ala Gly Asn Arg Val Arg Arg Ser Val Gly Ser Ser Leu181 185 190 195 200Ser Cys Ile Asn Leu Asp Trp Asp Val Ile Arg Asp Lys Thr Lys Thr Lys Ile Glu Ser201 205 210 215 220Leu Lys Glu His Gly Pro Ile Lys Asn Lys Met Ser Glu Ser Pro Asn Lys Thr Val Ser221 225 230 235 240Glu Glu Lys Ala Lys Gln Tyr Leu Glu Glu Phe His Gln Thr Ala Leu Glu His Pro Glu241 245 250 255 260Leu Ser Glu Leu Lys Thr Val Thr Gly Thr Asn Pro Val Phe Ala Gly Ala Asn Tyr Ala261 265 270 275 280Ala Trp Ala Val Asn Val Ala Gln Val Ile Asp Ser Glu Thr Ala Asp Asn Leu Glu Lys281 285 290 295 300Thr Thr Ala Ala Leu Ser Ile Leu Pro Gly Ile Gly Ser Val Met Gly Ile Ala Asp Gly301 305 310 315 320Ala Val His His Asn Thr Glu Glu Ile Val Ala Gln Ser Ile Ala Leu Ser Ser Leu Met321 325 330 335 340Val Ala Gln Ala Ile Pro Leu Val Gly Glu Leu Val Asp Ile Gly Phe Ala Ala Tyr Asn341 345 350 355 360Phe Val Glu Ser Ile Ile Asn Leu Phe Gln Val Val His Asn Ser Tyr Asn Arg Pro Ala361 365 370 375 380Tyr Ser Pro Gly His Lys Thr Gln Pro Phe Leu His Asp Gly Tyr Ala Val Ser Trp Asn381 385 390 395 400Thr Val Glu Asp Ser Ile Ile Arg Thr Gly Phe Gln Gly Glu Ser Gly His Asp Ile Lys401 405 410 415 420Ile Thr Ala Glu Asn Thr Pro Leu Pro Ile Ala Gly Val Leu Leu Pro Thr Ile Pro Gly421 425 430 435 440Lys Leu Asp Val Asn Lys Ser Lys Thr His Ile Ser Val Asn Gly Arg Lys Ile Arg Met441 445 450 455 460Arg Cys Arg Ala Ile Asp Gly Asp Val Thr Phe Cys Arg Pro Lys Ser Pro Val Tyr Val461 465 470 475 480Gly Asn Gly Val His Ala Asn Leu His Val Ala Phe His Arg Ser Ser Ser Glu Lys Ile481 485 490 495 500His Ser Asn Glu Ile Ser Ser Asp Ser Ile Gly Val Leu Gly Tyr Gln Lys Thr Val Asp501 505 510 515 520His Thr Lys Val Asn Ser Lys Leu Ser Leu Phe Phe Glu Ile Lys Ser521 525 530 535。