Table 1 Phage panning experiments ELISA plates (Dynatech Immulon I) were coated with BSA or (NPNA)3C-BSA and used in phage panning experiments. To the blocked antigen coated wells a total of 4 × 1010 pfu of the phage library in dilution buffer were added 1 × l010 pfu per well. After 4 h the wells were washed and phage eluted by applying either 0.1 M glycine-HCl pH 2.2 or a solution of the free peptide (~8 μM) (NPNA)3 dissolved in dilution buffer for 15 min at ambient temperature. An aliquot of the phage eluate was titered and the output determined.
Eluate after | Coating antigen / Elution method (×l05 pfu)* | |||
|---|---|---|---|---|
panning rounds | BSA / acid | BSA /(NPNA) 3 | (NPNA) 3 C BSA /acid | (NPNA) 3 C BSA /(NPNA) 3 |
1 | 2.9 (0.38) | 0.82 (0.032) | 3.0 (0.34) | 0.51 (0.024) |
2 | 1.4(0.03) | 0.24 (0.020) | 3.5 (0.24) | 1.5 (0.028) |
3 | 1.2(0.06) | 0.47 (0.020) | 4.3 (0.024) | 12 (0.68) |
4 | 13 (0.70) | 1.0(0.032) | 170 (30) | 370 (20) |