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    • 18. 发明授权
    • Preparation of chimaeric antibodies using the recombinant PCR strategy
    • 使用重组PCR策略制备嵌合抗体
    • US5858725A
    • 1999-01-12
    • US39198
    • 1993-07-29
    • James Scott CroweAlan Peter Lewis
    • James Scott CroweAlan Peter Lewis
    • C12N5/10C07K16/46C12N15/09C12N15/12C12P21/08C12Q1/68C12R1/91C07H21/04
    • C07K16/464C12Q1/686C07K2317/24C07K2319/00C07K2319/02Y10S530/867
    • The invention relates to a method of producing a chimaeric antibody in which the CDR of a first antibody is spliced between the framework regions of a second antibody. The method is performed using a template comprising two framework regions, AB and CD, and between them, the CDR which is to be replaced by a donor CDR. Primers A and B are used to amplify the framework region AB, and primers C and D used to amplify the framework region CD. However, the primers B and C also contain, at their 5' ends, additional sequence corresponding to all or at least part of the donor CDR sequence. Primers B and C overlap by a length sufficient to permit annealing of their 5' ends to each other under conditions which allow a polymerase chain reaction to be performed and thereby incorporate all of the donor CDR sequence. The amplified regions AB and CD may undergo splice overlap extension to produce the chimaeric product in a single reaction.
    • PCT No.PCT / GB91 / 01744 Sec。 371日期:1993年7月29日 102(e)日期1993年7月29日PCT 1990年10月8日PCT PCT。 出版物WO92 / 07075 日期1992年4月30日本发明涉及一种生产嵌合抗体的方法,其中第一抗体的CDR在第二抗体的框架区域之间被剪接。 该方法使用包含两个框架区AB和CD的模板进行,并且它们之间由供体CDR替代的CDR。 引物A和B用于扩增框架区AB,引物C和D用于扩增框架区CD。 然而,引物B和C在其5'末端还含有对应于全部或至少部分供体CDR序列的附加序列。 引物B和C的重叠长度足以使其5'末端在允许进行聚合酶链反应并因此并入所有供体CDR序列的条件下彼此退火。 扩增区域AB和CD可以经历拼接重叠延伸以在单个反应中产生嵌合产物。