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. 2011 Jan 1;3(1):49–60. doi: 10.4161/mabs.3.1.13989

Figure 3.

Figure 3

XOMA 052 epitope mapping. (A) The IL-1β PepSpot™ Peptide Array membrane probed with XOMA 052 reveals that XOMA 052 binds to peptide spots corresponding to amino acids 83–105 of the mature protein. (B) Alanine substituted peptides hybridized with XOMA 052. Sequences of the 16 peptides with the alanine substitution (in blue) are shown in the box below. Peptides 9–12 and 16 showed little or no binding by XOMA 052 (WT, wild type). (C) Sequence alignment of mature forms of mouse (m), human (h), rhesus (rh), rat (r) and rabbit (ra) IL-1β are shown. Residues that are identical in human, rhesus, rat and rabbit and differ in mouse are shown in bold and underlined. (D) Supernatants from wild type and six mutants of IL-1β (E64A, K65A, M95A, E96A, K97A and Q116E) were injected over XOMA 052 immobilized on a ProteOn XPR sensor chip. The fits of the off-rate data are shown as red lines. Mutants E96A, K97A and Q116E showed off-rates increased by 1,000-fold. (E) Sensorgrams of wild type and IL-1β mutants binding to sRI show that the mutant proteins were expressed and folded properly.