BDB - Library Details

Library

Library Name: SFTI-based phage display (SFTI8Ph) library (X8)

Library ID:

475

Library Length:

Library Complexity:

Library Titer:

Library From:

Uwe Haberkorn (German Cancer Research Center,(DKFZ) Heidelberg, Germany)

Library Randomness:

Completely random

Library Scheme:

NNN

Library Topology:

Linear

Comments:

The combinatorial sunflower trypsin inhibitor 1-based phage display (SFTI8Ph) library based on the sunflower trypsin inhibitor (SFTI) 1 scaffold structure was constructed by PCR. To this end, 8 amino acids except cysteine were randomly inserted between Thr4 and Cys11 in the binding loop of SFTI-1. The following oligonucleotides were used as templates for the PCR reaction: 5‘-TTACTCGCTCCATGGGCGGCAGGTGTACTNNN NNN NNN NNN NNN NNN NNN NNN TGTTATCCCGAT-3’ (SFTI8Ph forward; NNN = trimer encoding for one variable amino acid; Ella Biotech) and 5`-ATAATCTTGCGGCCGCACCGCCACCTGCTGCATCGGGATAACA-3` (SFTIPh reverse; Eurofins MWG Operon). With regard to the 125I-labeling of peptides the triplet TTT was exchanged by TAT to achieve a substitution of phenylalanine 12 to tyrosine. The PCR product was cloned in frame into the surface expression phagemid vector pSEX81 (Progen) and single clones were sequenced (GATC Biotech) to control the diversity of the library. For the preparation of phages presenting the peptides (theoretical diversity of 109) fused to the pIII coat protein on their surface the phagemid vector was transformed in XL1Blue bacteria.

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