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Figure 5 | BMC Microbiology

Figure 5

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 5

Competitive inhibition of PA(A)-PG-phages (A-D) and SpA-phage (E) binding to hIgG by PA(A)-PG combinations, PA(A)-PL combinations, SpA, SpG and both SpA plus SpG. 109TU of tested phages without and with each of 100 nM (black bars) or 25 nM (white bars) of inhibitor proteins were added into hIgG-coated wells respectively. Unbound phages were removed and 10 μl of 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, SpG and that both were used as inhibitors respectively.

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