Plasmid generation for TCR and antigen

TCR expression constructs were generated as previously described (21 (link)). HLA-C*07:01, HLA-C*07:02 cDNA, as well as truncated CSPG4 cDNA ending with codon 562 were cloned into pcDNA3.1/V5-His TOPO (Invitrogen, Waltham, MA, USA) using standard cloning techniques as previously described (22 (link)). Plasmids were propagated in 10-beta competent E. coli (NEB, Ipswich, MA, USA) and were purified using the Midi or Maxi Plasmid Prep kits from Qiagen (Hilden, Germany).

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Kropp K.N., Fatho M., Huduti E., Faust M., Lübcke S., Lennerz V., Paschen A., Theobald M., Wölfel T, & Wölfel C. (2023). Targeting the melanoma-associated antigen CSPG4 with HLA-C*07:01-restricted T-cell receptors. Frontiers in Immunology, 14, 1245559.

Publication 2023

pcDNA3.1/V5-His TOPO 10-beta competent E. coli

Cdna Codon Cspg4 E coli Hla c Plasmid Topo

Corresponding Organization : Johannes Gutenberg University Mainz

Other organizations : Heinrich Heine University Düsseldorf, Düsseldorf University Hospital

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Variable analysis

independent variables

HLA-C*07:01 cDNA
HLA-C*07:02 cDNA
Truncated CSPG4 cDNA ending with codon 562

dependent variables

Not explicitly mentioned

control variables

Not explicitly mentioned

controls

No positive or negative controls were explicitly mentioned.

Annotations

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