Covalent assembly of a soluble T cell receptor-peptide-major histocompatibility class I complex

被引:18
作者
Gregoire, C
Lin, SY
Mazza, G
Rebai, N
Luescher, IF
Malissen, B
机构
[1] CNRS MARSEILLE LUMINY, INSERM, CTR IMMUNOL, F-13288 MARSEILLE 9, FRANCE
[2] UNIV LAUSANNE, LUDWIG INST CANC RES, LAUSANNE BRANCH, CH-1066 EPALINGES, SWITZERLAND
关键词
D O I
10.1073/pnas.93.14.7184
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
We used stepwise photochemical crosslinking for specifically assembling soluble and covalent complexes made of a T-cell antigen receptor (TCR) and a class I molecule of the major histocompatibility complex (MHC) bound to an antigenic peptide, For that purpose, rye have produced in myeloma cells a single-chain Fv construct of a TCR specific for a photoreactive H-2K(d)-peptide complex. Photochemical cross-linking of this TCR single-chain Fv with a soluble form of the photoreactive H-2K(d)-peptide ligand resulted in the formation of a ternary covalent complex. We have characterized the soluble ternary complex and shoved that it reacted with antibodies specific for epitopes located either on the native TCR or on the K-d molecules, By preventing the fast dissociation kinetics observed with most T cell receptors, this approach provides a means of preparing soluble TCR-peptide-MHC complexes on large-scale levels.
引用
收藏
页码:7184 / 7189
页数:6
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