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Figure 6 | BMC Microbiology

Figure 6

From: Evolutional selection of a combinatorial phage library displaying randomly-rearranged various single domains of immunoglobulin (Ig)-binding proteins (IBPs) with four kinds of Ig molecules

Figure 6

Competitive inhibition of two representative PA(A)-PL-phages (A and B), SpA-phages (C) and 2L-phages (D) binding to hIgM molecules by PA(A)-PL combinations, PA(A)-PG combinations, SpA, 4L and both SpA plus 4L. 109TU of PA(A)-PL-phages, 1011TU of SpA-phages and 1010TU of 2L-phages without and with each of 100 nM (black bars) or 25 nM (white bars) of inhibitor proteins were added into hIgM-coated wells respectively. Unbound phages were removed and 10 μl exponentially growing E. coli TG1 was added into each well, incubated for 1 h at 37°C. The TG1 cells were harvested respectively and spread LB plates containing 100 μg/ml ampicillin, and bacterial colonies were counted after incubating at 37°C overnight. Inhibition rate was calculated: [1 - (mean of the bacterial colonies from tested wells with inhibitor proteins - mean of the bacterial colonies from blank control wells) divided by (mean of the bacterial colonies from tested wells without inhibitor proteins - mean of the bacterial colonies from blank control wells)] × 100%. No. 1 to 6: The expressed fusion proteins of PA(A)3N-PG-PA(A), PA(A)6N-PG, PA(A)-PG9N, PA(A)6N-PG3N, PA(A)9N-PL3N and PA(A)9N-PL were used as inhibitors respectively. No. 7 to 9: SpA, 4L and that both were used as inhibitors respectively.

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