Table 1 |
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|
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. |
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| Eluate after |
Coating antigen / Elution method (×l05 pfu)* |
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|
|
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| panning rounds |
BSA / acid |
BSA /(NPNA)3 |
(NPNA)3C BSA /acid |
(NPNA)3C BSA /(NPNA)3 |
|
|
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| 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) |
|
|
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|
* Figures represent the mean of the total plaque forming units eluted by either acid or excess free peptide, after repeated panning against BSA or (NPNA)3C-BSA. Values for the standard deviation are shown in brackets (). |
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|
Chappel et al. Malaria Journal 2004 3:28 doi:10.1186/1475-2875-3-28 |
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