- First Synthesis of each library is in one piece - peptide - sizes can be precisely predefined
- Very systematic peptide pattern designable without sequence preference to equal cost
- All sizes from smaller peptides to larger entities (Mini-)Proteins are possible (e.g. 8 – 40 aa)
- Defined peptides of high biological quality - no side reactions like in Peptide Chemistry
- 1:1 Stoichiometry of different Peptides can be guaranteed
- Smaller libraries size compared to random libraries with defined low complexity
- NO irrelevant Peptides NO empty vectors and comparable very low background in analysis
- NO loss of complexity through amplification
- Different carrier proteins (e.g. GST-tags) can flexible be applied to one and the same library
- Once synthesized the library can be propagated in a number of different forms!
- Deployment from Random Access Libraries to Epitope MicroArrays
- Ideal e.g. for AutoImmune Disease - AutoAntiBody – Individual Pattern Screenings
PepIDENT– libraries are very suitable for the analysis and the identification of specific protein-protein interactions in detail.
Dissecting proteins systematically into its minimal functional oligo-peptide entities for the identification of minimal specific binding sites is the strength of PepIDENT.
PepIDENT is designed in two different variants (1) epitope peptide random access library (PRAL) and (2) the MicroArray type of protein specific epitope libraries. The two types of library differ first in its design and second in its type of deployment.
Peptide epitope random access libraries use directly more or less complex mixtures of individually epitope-expressing E. coli clones which are just determined by the specificity of the epitope expressed.
These clones are plated randomly on the surface of a dish and subsequently the clones are transferred to a suitable reaction membrane. Before the antibody specific reaction identified the clones it is not clear which of the clone-specific expressed peptides are allocated to which antibody. This becomes clear by identifying the positive clones by an immunoreactions using antibodies and subsequently after sequencing the peptide coding DNA sequences of positive identified clones. Therefore access to the positive clones is random but specifically mediated by the epitope-paratope interaction.
First - for the most basic R&D work using the epitope peptide random access type of library these are a fast, reliable and comparably cheap way for epitope mappig and for getting valuable results.
But second - for example - patient related diagnostic tools based on PepIDENT libraries do have a higher need for fast and easy readout of the diagnostic information. In contrast to peptide random access libraries mentioned above the MicroArray based PepIDENT - technology provides additional positional information by selecting each of the possible clones in advance and positioning it on the MicroArray for allocating a positional code to it. This helps to identify relevant epitopes immediately without the need for sequencing.
In a third variant the PepIDENT libraries in its technical variant pEpMAP – PP is suitable for the analyses of specific interactions occurring between individual proteins specifically forming functional complexes with each other.
In addition in a forth variant (pEpMAP - Y2H) the peptide libraries created with PepIDENT can also be used for identifying specific interaction points in protein complexes in fusion with the yeast two hybrid system (Y2H) in order to gain complementary information about specific interaction mediating peptide sequences.
PepIDENT – technology forms the basis for the analysis of Epitope – Antibody - Paratope interactions and therefore establishes the basis for the analysis of molecular patho-pattern and related evidence based diagnostics to the relevant diseases.
In turn this generates the advantage to identify pattern of autoimmune reactions directed against known targeted proteins.
But in addition new target proteins can be identified in screening projects.
Especially for the characterization of autoimmune or allergen antibody pattern the identification of relevant epitopes is of great diagnostic value in case to case studies and provides the basis for the assessment of individualized therapeutic schemes.
Diagnostic devices for the identification of autoimmune- and allergene-antibodies, can be manufactured by use of the PepIDENT-technology.
This qualifies the PepIDENT – technology for having the potential to create products for the diagnostic market. To exploit its full potential these types of libraries need to be standardized and are replicated many times for placing it cost-effectively on the market.
Screening programs accompanying clinical trials or just point of care diagnostics close to the patient directly at the doctor's location e.g. the clinic can be performed with micro-array based libraries very easily.
Thus the PepIDENT - technology is powerful for both testing patients for patho-pattern and the development of therapeutic agents.