Table 1 Oligonucleotide primers used for construction of mutant libraries for affinity maturation of anti-CEA MA39 scFv antibody. Wobble symbol N means G or A or T or C, symbol S means G or C. In underlined part of oligonucleotides KM144 and KM145, each base was replaced with mixture of G/A/T/C at a frequency of 10%.

KM144

5'-TGTGCGAAAAGTAATGAGTTTCTTTTTGACTACTGGGGC-3'

KM145

5'-TCCGCCGAATACCACATAGGGCAACCACGGATAAGAGGAGTTACAGTAATAGTCAGCC-3'

KM149

5'-GTATATTACTGTGCGAAANNSNNSNNSAGTAATGAGTTTCTTNNSNNSNNSTTTGACTACTGGGGCCAG-3'

KM150

5'-GTATATTACTGTGCGAAANNSNNSAGTAATGAGTTTCTTNNSNNSNNSTTTGACTACTGGGGCCAG-3'

KM151

5'-GTATATTACTGTGCGAAANNSNNSNNSAGTAATGAGTTTCTTNNSNNSTTTGACTACTGGGGCCAG-3'

KM152

5'-GTATATTACTGTGCGAAANNSNNSAGTAATGAGTTTCTTNNSNNSTTTGACTACTGGGGCCAG-3'

KM148

5'-CTATTGCCTACGGCAGCCGCTGGA-3'

KM157

5'-TTTCGCACAGTAATATACGG-3'

KM158

5'-TATGTGGTATTCGGCGGA-3'

KM143

5'-GTCATCGTCGGAATCGTCATCTGC-3'

KM169

5'-GGTGGTAGCACATACTACGCA-3'

KM170

5'-GGAATTGTCTCTGGAGATGGTGA-3'

KM171

5'-TCGTCTGAGCTGACTCAGGAC-3'

KM172

5'-GCATGTGATCCTGACTGTCTG-3'

KM173

5'-CTCCAGCTCAGGAAACACAGCTT-3'

KM174

5'-AGCCTCATCTTCCGCCTGAG-3'

KM252

5'-GAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGC-3'

KM253

5'-GAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGC-3'