A single‐chain murine class I major transplantation antigen

Abstract

Single-chain mouse Kd molecules (SC-Kd) were engineered by connecting residue 276 of Kd heavy chain to the first residue of beta 2-microglobulin through spacers of various lengths, and expressed intracellularly in monkey COS-1 cells. Labeled SC-Kd molecules were found to react with several monoclonal antibodies which recognize native Kd molecules. SC-Kd-15 (with a spacer of 15 residues) was studied in more details. It could be purified and shown to regain a native-like structure after treatment with denaturing agents. Purified SC-Kd-15 could bind certain peptides in a manner qualitatively similar to the Kd.

Engineering and Characterization of a Stabilized α1/α2 Module of the Class I Major Histocompatibility Complex Product Ld

“…8). Early studies explored the linkage of ␤2m to the class I heavy chain (47) or of the peptide to class I heavy chain (48,49). It has been argued that single-chain strategies with peptides may be of variable success, due to the need for anchoring peptide termini into class I pockets.…”

Efficient binding to the MHC class I Kd molecule of synthetic peptides in which the anchoring position 2 does not fit the consensus motif

“…This peptide was previously shown to be Kd restricted [12]. The binding assays were performed with a single-chain, soluble H-2Kd molecule (SC-Kd) [16]. The C-terminal end of the cc3 domain of this molecule is connected to the N-terminal end of mouse pzmicroglobulin.…”

Recruitment of Oligoclonal Viral-Specific T cells to Kill Human Tumor Cells Using Single-Chain Antibody–Peptide–HLA Fusion Molecules

“…Therefore, the design of the CMV-scHLA-A2-SS1(scFv) fusion molecule as one single domain with a covalently linked peptide maximizes the stability of the peptide within the MHC groove, as it forces the peptide to be in close proximity with the MHC groove. Improving stability of the MHC molecule by covalently attaching the antigenic peptide was previously demonstrated in our laboratory and by others who used soluble MHC molecules with covalently bound peptides to study the structure and stability of T-cell receptor (TCR)-MHC interactions (17,(35)(36)(37). For the CMV-scHLA-A2-SS1(scFv) fusion molecules, overhang of the linker from the MHC groove was tolerated and no changes in TCR recognition were observed.…”