Title:
組換えＰＣＲ法を用いるキメラ抗体の調製
Document Type and Number:
Japanese Patent JP3516950
Kind Code:
B2
Abstract:
PCT No. PCT/GB91/01744 Sec. 371 Date Jul. 29, 1993 Sec. 102(e) Date Jul. 29, 1993 PCT Filed Oct. 8, 1990 PCT Pub. No. WO92/07075 PCT Pub. Date Apr. 30, 1992The 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.
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Inventors:
クロウ、ジェイムズ・スコット
ルウイス、アラン・ピーター
ルウイス、アラン・ピーター
Application Number:
JP51645691A
Publication Date:
April 05, 2004
Filing Date:
October 08, 1991
Export Citation:
Assignee:
ザ・ウエルカム・ファウンデーション・リミテッド
International Classes:
C12N5/10; C07K16/46; C12N15/09; C12N15/12; C12P21/08; C12Q1/68; C12Q1/686; C12R1/91; (IPC1-7): C12N15/09; C12P21/08
Other References:
Ｒ．Ｍ．ＨＯＲＴＯＮ，Ｅｎｇｉｎｅｅｒｉｎｇ ｈｙｂｒｉｄ ｇｅｎｅｓ ｗｉｔｈｏｕｔ ｔｈｅ ｕｓｅ ｏｆ ｒｅｓｔｒｉｃｔｉｏｎ ｅｎｚｙｍｅｓ：ｇｅｎｅ ｓｐｌｉｃｉｎｇ ｂｙ ｏｖｅｒｌａｐ ｅｘｔｅｎｓｉｏｎ，Ｇｅｎｅ（１９８９），ＮＬ，Ｖｏｌ．７７，Ｎｏ．１，ｐ．６１−６８
Ｒ．Ｍ．ＨＯＲＴＯＮ，Ｇｅｎｅ Ｓｐｌｉｃｉｎｇ ｂｙ Ｏｖｅｒｌａｐ Ｅｘｔｅｎｔｉｏｎ：Ｔａｉｌｏｒ−Ｍａｄｅ Ｇｅｎｅｓ Ｕｓｉｎｇ ｔｈｅ Ｐｏｌｙｍｅｒａｓｅ Ｃｈａｉｎ Ｒｅａｃｔｉｏｎ，Ｂｉｏｔｅｃｈｎｉｑｕｅｓ（１９９０−Ｍａｙ），米国，Ｖｏｌ．８，Ｎｏ．５，ｐ．５２８−５３５
Ｒ．Ｍ．ＨＯＲＴＯＮ，Ｇｅｎｅ Ｓｐｌｉｃｉｎｇ ｂｙ Ｏｖｅｒｌａｐ Ｅｘｔｅｎｔｉｏｎ：Ｔａｉｌｏｒ−Ｍａｄｅ Ｇｅｎｅｓ Ｕｓｉｎｇ ｔｈｅ Ｐｏｌｙｍｅｒａｓｅ Ｃｈａｉｎ Ｒｅａｃｔｉｏｎ，Ｂｉｏｔｅｃｈｎｉｑｕｅｓ（１９９０−Ｍａｙ），米国，Ｖｏｌ．８，Ｎｏ．５，ｐ．５２８−５３５
Attorney, Agent or Firm:
鈴江 武彦 （外３名）