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Representative T cell hybridomas were isolated after HEL (11–27) or HEL (11–27⁹ᴱ) immunizations (clones 21.30.7 and 72.10, respectively) and tested for activation by HEL (11–27) and mutant peptides. Clone 21.30.7 (also called 21.30) was, analogous to the polyclonal response, strongly influenced by the presence of a negative charge at P9 unlike clone 72.10. TCR 21.30 was approximately 100 times more sensitive to HEL (11–27) than HEL (11–27⁹ᴱ). The TCR α and β chains of these clones were isolated by RT-PCR, sequenced, and used to produce recombinant soluble TCR molecules suitable for structural studies. It was noteworthy that TCR 21.30 had a negatively charged residue (Aspβ109) at position 3 of its CDR3β loop (WDRAGNTL).
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