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Figure 1.
Figure 1. Isolation of MAb5G8-specific V[NAR] domains. (A)
Pools of polyclonal IgNAR-bacteriophages displaying V[NAR]
domains were assayed for binding to MAb5G8 (selection antigen)
or 2% MPBS (nonspecific control) pre-pan and following one to
four rounds of library panning. Bound bacteriophages were
detected using an HRP conjugated anti-fd phage antibody, and
results represent the average of triplicate wells. The positive
control represents bacteriophages displaying the MAb5G8-specific
E2 peptide [^NEDENTLQHAYPID^C]. (B) Protein alignment of deduced
amino acid sequences for V[NAR]s 1-A-2, 1-A-7, 1-A-11, and
1-A-14. CDR1 and CDR3 regions are highlighted and identical
residues (dark shading) and conservative replacements (light
shading; I/V/L/M, D/E, K/R, A/G, T/S, Q/N, F/Y) indicated. (C)
Expanded comparison of V[NAR] 1-A-2, 1-A-7, 1-A-11, and 1-A-14
CDR regions. Potential peptide motifs for interaction with
MAb5G8 are underlined, and 1-A-11 cysteine residues positioned
to form an inter-loop disulphide linkage are highlighted in bold.
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