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Cloning and Characterisation of TPO Autoantibodies Using Combinatorial Phage Display Libraries

 

作者: HexhamM. J.,   PartridgeJ. I.,   FurmaniakJ.,   PetersenB. V.,   CollsC. J.,   PeggC.,   SmithRees B.,   BurtonR. D.,  

 

期刊: Autoimmunity  (Taylor Available online 1994)
卷期: Volume 17, issue 3  

页码: 167-179

 

ISSN:0891-6934

 

年代: 1994

 

DOI:10.3109/08916939409010651

 

出版商: Taylor&Francis

 

关键词: Thyroid;Autoantibody;Thyroid Peroxidase;Antibody Repertoire;Hashimoto's disease

 

数据来源: Taylor

 

摘要:

Thyroid lymphocyte RNA from a Hashimoto patient with high serum levels of autoantibodies to thyroid peroxidase (TPO) was used to construct a phage display antibody library in the phagemid vector pComb3. The library (100,000cfu) encoded IgG 1 heavy chains together with kappa light chains. Selection of the phages displaying TPO antibody on TPO-coated ELISA plates yielded a phage population enriched for surface expression of TPO antibody Fabs. 3 different Fabs specific for TPO were subsequently isolated with affinities in the region of 109molar”’. 2 of the Fabs recognised the same, or closely related, epitopes on TPO whereas the third Fab recognised a different epitope. These 2 epitopes were recognised by TPO autoantibodies in the serum of the lymphocyte donor and a series of 10 patient sera. Available sequence data showed that several non-self antibodies and non-thyroid autoantibodies use the same V kappa and VHgermline genes as TPO autoantibodies. There appeared to be no clear relationship between gene sequence or gene family usage by TPO autoantibodies of the same or similar epitope specificity.

 

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