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Title 

Generation of antibodies recognizing an aberrant glycoform of human tissue inhibitor of metalloproteinase-1 (TIMP-1) using decoy immunization and phage display

 

디코이면역법과 파아지 디스플레이를 이용한 TIMP-1 단백질의 당변이형을 인식하는 항체 제조

Authors 

Hyun Ju AnYong Sam KimChul Ho LeeEun Wie ChoHyang Sook YooSeung Ho KimJeong Heon KoSang Jick Kim

Publisher 

Elsevier

Issue Date 

2011

Citation 

Journal of Biotechnology, vol. 151, no. 2, pp. 225-230

Keywords 

Antibody libraryDecoy immunizationGlycanN-linked β(1,6)-GlcNAc branchPhage display

Abstract 

Aberrant glycosylation of human tissue inhibitor of metalloproteinase-1 (TIMP-1) by N-acetylglucosaminyltransferase-V (GnT-V) was previously reported to be related to cancer progression. Here, we report on the antibodies recognizing the structural features initiated by an addition of N-linked β(1,6)- N-acetylglucosamine (GlcNAc) branch by GnT-V on TIMP-1. Two glycoforms of TIMP-1, TIMP1-L produced in Lec4 cells without GnT-V activity and TIMP1-B in GnT-V overexpressing transfectant cells, were purified from culture supernatant and used for generation of antibodies. TIMP1-L was injected in the left hind footpad of mice as decoy and TIMP1-B in the right hind footpad as immunogen. Phage-displayed scFv library was constructed from the B cells retrieved from the right popliteal lymph nodes and subjected to panning and screening. Phage ELISA of individual clones revealed the scFv clones with preferential binding activity to TIMP1-B, and they were converted into an scFv-Fc format for further characterization of binding specificity. Glycan binding assay of an antibody, 1-9F, revealed its differential specificity toward an extension of glycan structure initiated with β(1,6)-GlcNAc linkage and terminally decorated with a sialic acid. This study demonstrates feasibility of a new strategy combining decoy immunization with phage display for the efficient generation of antibodies tracking down structural features of different glycoforms.

ISSN 

0168-1656

Link 

http://dx.doi.org/10.1016/j.jbiotec.2010.12.004

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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