[发明专利]人甲状旁腺激素(1-34)二联体与人血清白蛋白的融合蛋白及其制备

摘要

人甲状旁腺激素(1-34)(hPTH(1-34))二联体与人血清白蛋白(HSA)的融合蛋白及其制备技术的发明属于长效重组融合蛋白技术领域。本发明的融合蛋白包括与人甲状旁腺激素(1-34)至少85％序列同源的二联体的第一区和与人血清白蛋白至少85％序列同源的第二区。利用两次重叠PCR技术，现将两个hPTH(1-34)进行拼接得到二联体，再将二联体与HSA拼接，中间不加入任何连接肽。该融合蛋白可以在不改变融合蛋白特性的前提下，进行个别氨基酸残基的取代、缺失或加入，它们在保留了人甲状旁腺激素激活受体和刺激骨重建的作用的基础上，显著延长了在体内的半衰期，是药学领域中，能够治疗骨质疏松症的，有良好前景的药用融合蛋白。

主权项

1、人甲状旁腺激素(1-34)二联体与人血清白蛋白的融合蛋白的基因序列和蛋白质序列，如下：(1)基因序列：TCCGTTTCCGAGATTCAATTGATGCACAACTTGGGCAAGCACTTGAACTCCATGGAGAGAGTTGAGTGGTTGAGAAAGAAGTTGCAAGACGTTCACAACTTCTCCGTTTCCGAGATTCAATTGATGCACAACTTGGGCAAGCACTTGAACTCCATGGAGAGAGTTGAGTGGTTGAGAAAGAAGTTGCAAGACGTTCACAACTTCGATGCACACAAGAGTGAGGTTGCTCATCGGTTTAAAGATTTGGGAGAAGAAAATTTCAAAGCCTTGGTGTTGATTGCCTTTGCTCAGTATCTTCAGCAGTGTCCATTTGAAGATCATGTAAAATTAGTGAATGAAGTAACTGAATTTGCAAAAACATGTGTTGCTGATGAGTCAGCTGAAAATTGTGACAAATCACTTCATACCCTTTTTGGAGACAAATTATGCACAGTTGCAACTCTTCGTGAAACCTATGGTGAAATGGCTGACTGCTGTGCAAAACAAGAACCTGAGAGAAATGAATGCTTCTTGCAACACAAAGATGACAACCCAAACCTCCCCCGATTGGTGAGACCAGAGGTTGATGTGATGTGCACTGCTTTTCATGACAATGAAGAGACATTTTTGAAAAAATACTTATATGAAATTGCCAGAAGACATCCTTACTTTTATGCCCCGGAACTCCTTTTCTTTGCTAAAAGGTATAAAGCTGCTTTTACAGAATGTTGCCAAGCTGCTGATAAAGCTGCCTGCCTGTTGCCAAAGCTCGATGAACTTCGGGATGAAGGGAAGGCTTCGTCTGCCAAACAGAGACTCAAGTGTGCCAGTCTCCAAAAATTTGGAGAAAGAGCTTTCAAAGCATGGGCAGTAGCTCGCCTGAGCCAGAGATTTCCCAAAGCTGAGTTTGCAGAAGTTTCCAAGTTAGTGACAGATCTTACCAAAGTCCACACGGAATGCTGCCATGGAGATCTGCTTGAATGTGCTGATGACAGGGCGGACCTTGCCAAGTATATCTGTGAAAATCAAGATTCGATCTCCAGTAAACTGAAGGAATGCTGTGAAAAACCTCTGTTGGAAAAATCCCACTGCATTGCCGAAGTGGAAAATGATGAGATGCCTGCTGACTTGCCTTCATTAGCTGCTGATTTTGTTGAAAGTAAGGATGTTTGCAAAAACTATGCTGAGGCAAAGGATGTCTTCCTGGGCATGTTTTTGTATGAATATGCAAGAAGGCATCCTGATTACTCTGTCGTGCTGCTGCTGAGACTTGCCAAGACATATGAAACCACTCTAGAGAAGTGCTGTGCCGCTGCAGATCCTCATGAATGCTATGCCAAAGTGTTCGATGAATTTAAACCTCTTGTGGAAGAGCCTCAGAATTTAATCAAACAAAATTGTGAGCTTTTTGAGCAGCTTGGAGAGTACAAATTCCAGAATGCGCTATTAGTTCGTTACACCAAGAAAGTACCCCAAGTGTCAACTCCAACTCTTGTAGAGGTCTCAAGAAACCTAGGAAAAGTGGGCAGCAAATGTTGTAAACATCCTGAAGCAAAAAGAATGCCCTGTGCAGAAGACTATCTATCCGTGGTCCTGAACCAGTTATGTGTGTTGCATGAGAAAACGCCAGTAAGTGACAGAGTCACCAAATGCTGCACAGAATCCTTGGTGAACAGGCGACCATGCTTTTCAGCTCTGGAAGTCGATGAAACATACGTTCCCAAAGAGTTTAATGCTGAAACATTCACCTTCCATGCAGATATATGCACACTTTCTGAGAAGGAGAGACAAATCAAGAAACAAACTGCACTTGTTGAGCTCGTGAAACACAAGCCCAAGGCAACAAAAGAGCAACTGAAAGCTGTTATGGATGATTTCGCAGCTTTTGTAGAGAAGTGCTGCAAGGCTGACGATAAGGAGACCTGCTTTGCCGAGGAGGGTAAAAAACTTGTTGCTGCAAGTCAAGCTGCCTTAGGCTTA(2)氨基酸序列：SVSEIQLMHN LGKHLNSMER VEWLRKKLQD VHNFSVSEIQ LMHNLGKHLN SMERVEWLRKKLQDVHNFDA HKSEVAHRFK DLGEENFKAL VLIAFAQYLQ QCPFEDHVKL VNEVTEFAKTCVADESAENC DKSLHTLFGD KLCTVATLRE TYGEMADCCA KQEPERNECF LQHKDDNPNLPRLVRPEVDV MCTAFHDNEE TFLKKYLYEI ARRHPYFYAP ELLFFAKRYK AAFTECCQAADKAACLLPKL DELRDEGKAS SAKQRLKCAS LQKFGERAFK AWAVARLSQR FPKAEFAEVSKLVTDLTKVH TECCHGDLLE CADDRADLAK YICENQDSIS SKLKECCEKP LLEKSHCIAEVENDEMPADL PSLAADFVES KDVCKNYAEA KDVFLGMFLY EYARRHPDYS VVLLLRLAKTYETTLEKCCA AADPHECYAK VFDEFKPLVE EPQNLIKQNC ELFEQLGEYK FQNALLVRYTKKVPQVSTPT LVEVSRNLGK VGSKCCKHPE AKRMPCAEDY LSVVLNQLCV LHEKTPVSDRVTKCCTESLV NRRPCFSALE VDETYVPKEF NAETFTFHAD ICTLSEKERQ IKKQTALVELVKHKPKATKE QLKAVMDDFA AFVEKCCKAD DKETCFAEEG KKLVAASQAA LGL