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V1.2-100702 Version PepID - bioPeptide Library – Self-Cloning Kit: MultiPepbio Expression, Detection, Identification and Validation of Minimal Antigens & Epitopes For your specific PepID - online designs and order of your PepID-library please use: https://www.atg-biosynthetics.com/index.php?page=library-design The PepID - methodological work flow. Digital protein or gene sequences available from online data bases are dissected into sub-sequences in variable designs. Genes are first reverse translated into individual coding sequences (CDS). The individual bio-peptide CDS designs are then realized by de novo double stranded DNA synthesis and then physically implemented in a compressed form in a CDS backup vector pPepCDS. From this all coding sequences can be reproduced infinitely always in a 1:1 stoichometry. The pPepCDS coding sequence individual bio-peptide entities are then cloned into the library expression vector pEPX1. The vector pEPX1 is used for expression cloning of bio-peptides fused to a variable carrier protein free of choice for its use in the bioAssay systems they are designed for. ATG:biosynthetics GmbH · Weberstr. 40 D-79249 Merzhausen · Germany · P +49 (0)761-8889424 F +49 (0)761-8889425 M +49 (0)179-6607141 VOIP-Access at Skype : atgbio [email protected] e-mail www.ATG-Biosynthetics.com www -1- · V1.2-100702 Version Advantages of the PepID - bioPeptide Library Concept and the Advantages of the Ribosomal Synthesis Relative to Chemical Peptide Synthesis First Synthesis of each library is in one piece - biopeptides can be released, stored and reproduced infinitely on demand in always the same quality 1:1 Stoichiometry of different bioPeptides of the library can be guaranteed Flexible, variable and systematic peptide pattern sizes can be precisely predefined and are free designable without sequence preference of synthesis conditions to equal cost All sizes from smaller peptides to larger entities (Mini-)Proteins are possible (e.g. 8 – 50+ aa) Defined peptides of very high biological quality - no extensive side reactions like in peptide chemistry NO loss of complexity through amplification – compared to random libraries – Phage Display — constant consistency of library quality can be guaranteed Compared to phage display random libraries dramatically smaller libraries size with defined low complexity — semirational approach NO irrelevant bioPeptides - NO empty vectors comparable very low background in analysis Modular Design - different carrier proteins (e.g. GST-tags) flexible applied to one and the same library. Once synthesized the library can be propagated in a number of different forms — flexible change of carrier and reporter genes is possible. Deployment from simplest Random Access Screening Library Platings to focussed Epitope MicroArrays to production of higher quantities for Therapeutic deployment Ideal for individual patient epitope pattern screenings e.g AutoImmune Disease – AutoAntiBody PepID-Vectors: pPepCDS are the Library Backup/ Source Vectors and pEPX1 different Library expression vectors. Each of the individual peptide-coding sequences (PepCDS) are coding for one of the designed peptides. Thus the pPepCDS vectors contain all single bio-peptide-CDSs (X) of a protein peptide library design in a compact formate in pPepCDS. These can be released and each of it is cloned individually by fusing it to a custom carrier protein of desired properties like the GSTTag (Gluthatione-S-Transferase) for it‘s expression in pEPX1-X. Different carrier proteins are available either with an affinity tag or as a reporter tag. Bio-peptide libraries can be self-cloned or provided custom made for any individual protein sequence desired which is available (e.g. from digital data bases). Compared to the classical DNaseI—fragment libraries frequently prepared for the generation of non-random libraries PepID-Libraries use only peptide CDS cloned in frame! Achieve a Maximum of Robustness by Simplification of Proven Procedures - Synthetic Genes for Peptide Synthesis - -2-