PepIDENT Inquiry

PepIDENT
bioPeptides

PepID- Epitope - Screening/ Mining - Libraries


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Peptide Expression Libraries in two different MODES:

MODE I: SINGLE-Library - MODE - Type A and Type B
DNA -Coding Peptide Sequences of your Library are fused in frame to the carrier protein of your choice.
MODE I+II: DUAL- Library - MODE - Type A and Type B
DNA -Coding Peptide Sequences of your Library are fused in frame to the reporter gene of your choice.
For Antibody binding analysis typically 33% to 66% overlaps are used. You can also use your specific strategy for designing the systematic peptide sequence pattern of your desire.
MODE_I: SINGLE-Library - MODE: A Primary Screening - Peptide Library is Fused to specific Peptide Carrier Genes

Type-A:
RANDOM ACCESS - Peptide Expression Library - if you are screening for the best mode of function your specific bioAssay allows you to identify the clones which show the functional properties you are looking for. Clones with the best mode of function can be sequenced to identify the function allocated sequence.
Please select the Carrier-Genes/Proteins of your choice:
Yeast-Gal4
Gluthatione- S -Transferase-Tag SJ26-GST
gIII - Protein
gVIII - Protein
His-Tag
Streptavidine
ProteinA
CTB (Cholera Toxin B sub unit)
MicroBody-Genes
Type-B:
Mode II - Type B: MICRO ARRAY - Peptide Library - this type of Peptide Library is positional preselected in that each position resembles one specific peptide clone which does not need to be identified.
MODE_I+II: DUAL - Library - MODE: An additional Second Screening - Peptide Library is Fused to specific Reporter Gens

Reporter-Genes/Proteins:
Gluthatione- S -Transferase-Tag SJ26-GST
CAT - Chloramphenicol - Acetyl-Transferase
eGFP - Protein (green fluorescence protein)
RFP - Protein (red fluorescence protein)
Luciferase - gene
GUS - Glucuronidase
LacZ - β -Galactosidase
ATG delivers your peptide expression library which is manufactured according to your peptide pattern strategy or idea.
Option 1: You provide ATG with your specific strategy ... Library Design Tool ...
Option 2: Use ATG service to develop your Peptide Library Strategy ... please inquire here ...
This starts with literature research and project planning. Comprehensive data mining of publications of the available literature related to your protein for known function its presumable interaction partners and sequence analyses. Detailed planning of the specific interaction mapping peptide pattern designs and the experimental design on the basis of the informational research results is performed. The experimental design strategy is provided and finally according to the specific strategy of systematic peptide sequence pattern are derived. Results are provided to you for your control.
Library DNA syntheses by ATG
  • Back-translation of the amino acid sequences.
  • Gene synthesis in basic standard vector like pUC18 is performed at ATG. First the library is generated in an un-expressed state in one piece. So we can guarantee its completeness and document it.
  • Sequence documentation is provided for a final control prior and after to synthesis.
Expression cloning by ATG
The carrier fusion-protein - bound peptide library is generated (f) E. coli T7 expression system is envisioned. Molecular cloning of the peptide library coding sequences (CDS) into a pEpMAP - type vector system is performed. Consequently, genes will be synthesized optimized for high E. coli expression (e.g. strong Shine Dalgarno sequence, optimal codon usage) and cloned in ATG proprietary vector with an Arabinose regulated promoter Bacterial strains like BL21, Tuner or Origami show enhanced solubility properties of proteins and/or enhanced cystein disulfide-bridge formation for improving specific activity. Consequently strains will be tested comparatively for expression finally to be determined.
( http://www.emdbiosciences.com/html/NVG/solubility.html )
PepID and Epitope - Mapping or Protein Interaction Screening by ATG
Epitope - mAB, nanoBody, Affibody, domainAB, protein-protein interactions are screenend by ATG

Final Documentation - Analysis Data are provided by ATG

The final documentation is provided with SOPs for all procedures developed and in addition all real and relevant data measured are provided according to ISO standards and in the metric system. Purchase management, specific use of equipment used and chemicals.

Note :
We assure that in passing our quality management the libraries are shipped out from us correctly and according to your functional and quality requirement specifications.
Please kindly verify the properties and confirm it with us as soon as possible upon receipt of the product we delivered to you.
For safety reasons in order to avoid any loss of your property (e.g.during transport), we will keep a backup of the SOP relevant equipment for one month.
During this period, please contact us promptly in case if you have any problem so that we can re-check it.
This is the time frame you can apply if your material is lost or erroneous for product liability issues.

Confidentiality Statement:
ATG:biosynthetics declares that we will keep your intellectual property about your genes/ protein sequences and protocols in strict confidentiality and we will NOT, in any manner, disclose, develop and utilize your genes/sequences. We will destroy all the related testing data and results after one month.