Three-Step Seromarker Discovery Service

JPT Peptide Technologies is now able to offer a modular three-step assay service for biomarker screening and biomarker discovery in patient samples. The antibody epitope and seromarker profiling service workflow analyzes the humoral immune response in three steps. The three assay service modules can be combined or utilized individually according to your requirements.

  1. High throughput biomarker screening (e.g. seromarker screening), using thousands of custom peptides
  2. Verification of the peptide hits for hundreds of samples
  3. Validation of results by robust custom peptide ELISA

Our service includes:

  • Experienced scientists work with you on the optimal biomarker screening strategy for your biomarker discovery project
  • We help to design the peptide sequences
  • Production of tailored PepStar™ peptide microarrays and/or custom peptide ELISA plates
  • Optimization of arrays and peptides for your requirements
  • Biomarker screening and control experiments using your samples
  • Peptide screening platform compatible with antibodies, sera, whole blood and other biological fluids that contain antibodies
  • JPT's biologists and computer scientists perform data evaluation and analysis
  • Experiments and results are delivered as a detailed report

Multiplexed Epitope Discovery

Assay Format: High content peptide microarray
No. of Peptides: Up to 6912 peptides in triplicates per sample
No. of Samples
: 1 sample per slide
Principle:
Large number of peptides tested against a limited number of samples
Advantages:
Low cost per peptide, high peptide throughput
Application:
First screening round for the selection of relevant peptides from thousands of candidate peptides

Selective Antigen Profiling

Assay Format: Multiwell peptide microarray
No. of Peptides: Up to 188 peptides in triplicates per sample
No. of Samples:
21 samples per slide
Principle:
Selected peptides tested against a large number of samples
Advantages:
Low cost per sample, high sample throughput
Application:
Second screening round to verify candidate peptides for a significant number of samples

Marker Validation

Assay Format: Peptide ELISA
No. of Peptides: Up to 96 peptides per plate
No. of Samples:
Up to 96 samples per plate
Principle:
Flexible for low numbers of peptides & samples
Advantages:
Economic common assay
Application:
Third screening round for validation of hits or diagnostic assays

Case Study: Epitope Mapping of Epstein-Barr Virus Antigens

High density peptide microarrays have been proven to be efficient tools to map the immune response at a single epitope resolution for large panels of antigens requiring only minute amount of serum or other patient samples (1 µl per assay) to perform a complete analysis [Barouch et al., 2013].

Herein we present a comprehensive but modular humoral immune response profiling platform allowing a systematic and high resolution analysis of the individual antibody repertoire comprising the following techniques:
(1) Multiplexed Epitope Discovery: a limited number of samples (typically 20 to 200) are profiled on PepStar™ High Content Peptide Microarrays displaying up to 6912 peptides in triplicates. This allows the coverage of a broad range of known and potentially relevant cancer antigens as well as antigens from infectious agents.
(2) Selective Antigen Profiling: pre-selected antigens and peptides will be tested against a large number of samples (typically 200 to 1000). For this purpose PepStar™ Multiwell Peptide Microarrays will be used taking advantage of parallel and economic testing of 21 patient samples on a single microarray. The format is limited to 188 candidate peptides (in triplicate) per sample allowing systematic profiling of candidate peptides with large numbers of test samples.
(3) Marker Validation: a robust Peptide ELISA platform was developed based on highly purified peptides allowing both, the validation of the selected candidates as well as detailed analysis of the recognition. The platform enables transfer of results towards diagnostic assays and the selection of vaccine candidates.

The performance of this new platform is exemplarily demonstrated for selecting relevant epitopes from a large panel of antigens of the Epstein-Barr Virus.

Read more in our Application Note.

Selected References for Three-Step Seromarker Profiling


"Macrophage Migration Inhibitory Factor-CXCR4 Receptor Interactions: Evidence for Partial Allosteric Agonism in Comparison with CXCL 12 Chemokine"
Rajasekaran et al., J Biol Chem. (2016) - PMID: 27226569
"Antibodies Reacting with JCPyV_VP2 _167-15mer as a Novel Serological Marker for JC Polyomavirus Infection"
Lagatie et al., Virology Journal (2014) - PMID: 25273879

Read More References

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Three-Step Seromarker Profiling

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