Use of SpyTag/SpyCatcher to construct bispecific antibodies that target two epitopes of a single antigen

被引：26

作者：

Yumura, Kyohei ^{[1
]}

Akiba, Hiroki ^{[2
]}

Nagatoishi, Satoru ^{[2
]}

Kusano-Arai, Osamu ^{[3
]}

Iwanari, Hiroko ^{[3
]}

Hamakubo, Takao ^{[3
]}

Tsumoto, Kouhei ^{[1
,2
,4
]}

机构：

[1] Univ Tokyo, Grad Sch Frontier Sci, Dept Med Genome Sci, Kashiwa, Chiba 2778562, Japan

[2] Univ Tokyo, Sch Engn, Dept Bioengn, Bunkyo Ku, Tokyo 1138656, Japan

[3] Univ Tokyo, Dept Quantitat Biol & Med, Res Ctr Adv Sci & Technol, Tokyo 1538904, Japan

[4] Univ Tokyo, Inst Med Sci, Med Proteom Lab, Minato Ku, Tokyo 1088639, Japan

来源：

JOURNAL OF BIOCHEMISTRY | 2017年 / 162卷 / 03期

基金：

日本学术振兴会; 日本科学技术振兴机构;

关键词：

biolayer interferometry; bispecific antibody; isothermal titration calorimetry; single-chain Fv; SpyTag; ANTITUMOR-ACTIVITY; PROTEIN; FRAGMENTS; BACULOVIRUS; AFFINITY; ROBO1;

D O I：

10.1093/jb/mvx023

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Bispecific antibody targeting of two different antigens is promising, but when fragment-based antibodies are used, homogeneous production is difficult. To overcome this difficulty, we developed a method using the SpyTag/SpyCatcher system in which a covalent bond is formed between the two polypeptides. Using this method, we constructed a bispecific antibody that simultaneously interacted with two different epitopes of roundabout homologue 1 (ROBO1), a membrane protein associated with cancer progression. A bispecific tetravalent antibody with an additional functional moiety was also constructed by using a dimeric biotin-binding protein. An interaction analysis of ROBO1-expressing cells and the recombinant antigen demonstrated the improved binding ability of the bispecific antibodies through spontaneous binding of the two antibody fragments to their respective epitopes. In addition, multivalency delayed dissociation, which is advantageous in therapy and diagnosis.

引用

页码：203 / 210

页数：8

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