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

Abstract:

: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.

journal_name

J Biochem

journal_title

Journal of biochemistry

authors

Yumura K,Akiba H,Nagatoishi S,Kusano-Arai O,Iwanari H,Hamakubo T,Tsumoto K

doi

10.1093/jb/mvx023

subject

Has Abstract

pub_date

2017-09-01 00:00:00

pages

203-210

issue

3

eissn

0021-924X

issn

1756-2651

pii

3869163

journal_volume

162

pub_type

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