HLA Peptide Target Reagents
to develop T cell epitope-based Immunotherapies.
HLA Peptide Complex Engineering
HLA Peptide Target Reagents
Using soluble HLA technology, we offer small and large-scale manufacturing of peptide-modified HLA complexes harboring a specific peptide of your choice used as reagents for Immuno-Therapeutic and Diagnostic HLA Receptor Generation and Validation.
How it Works
Selection of Peptide, sHLA Molecule and Modality
To start the custom manufacturing of a specific HLA peptide complex, select a target peptide, a sHLA allele and the modality for the assembly. HLA peptide epitopes must be carefully selected. Successful candidates mostly derive from target discovery pipelines showing high promise in being immunologically recognized by T cells in the fight against cancer and infectious diseases. A broad representation of HLA alleles is available to select from at high purity levels to guarantee outstanding performance. Modalities are selected based on the application intended and can be a simple monomer, a biotinylated complex or a tetramer.
Validation of Target Peptide
Before initiating an exchange process, it is important to validate individual target peptides for HLA binding. For each project peptide, inhibitory concentrations are determined by plotting the response values as a function of the logarithms of competitor concentrations and applying a nonlinear regression dose response model with variable slope for analysis. The calculated logIC50 value is reported as measure of the effectiveness of the target peptide.
Execution of Peptide Exchange Process
Depending on the product modality, distinct peptide exchange process pipelines have been developed following multi-step protocols that include loading, peptide recovery, purification, enrichment, quantitation, and quality control stages. Each process addresses multiple aspects of increased purity by removing excess assembly components that potentially cause downstream interferences. The assembly is initiated by using a pre-manufactured soluble HLA molecule. All processes deliver stable products in a highly reproducible fashion.
Integrity and Performance Qualification
For validation of peptide exchanged monomers and tetramers, a selective, physiologically relevant assay will be applied to report on the reagent’s potency and stability, providing an assessment of the molecule’s biological activity. Confirming proper structural configuration is a major part of our quality control validation procedure using best analytical practices. Assays involve the classical W6/32 – Anti-b2m Sandwich ELISA Format and dose-response curve assessments providing proper validation on the function and behavior of HLA molecules where structurally intact HLA proteins are critical.
Mass-Spec Exchange Confirmation
Unprecedented, Pure Protein is the only company currently offering nano-LC Mass-Spec analysis to provide confirmation that the peptide of interest is solely presented by the engineered HLA complex. After denaturation of a small amount of engineered sHLA, ion chromatograms for the peptide of interest are extracted and compared to the chromatogram from the native HLA molecule. The process can achieve up to 99.9% exchange efficiency.
High Quality Molecules
Naturally folded soluble HLA molecules produced in human cell lines (not bacterial), at a purity level >95% using affinity chromatography.
Choose from a wide range of major HLA alleles stably expressed in mammalian cell lines and available in small and large quantities.
All cell lines are confirmed by sequence-based typing, the manufacturing process traced using activity measurements and each production validated.
HLA peptide complexes are available in different formats including non-modified monomers, biotinylated monomers and non-labeled tetramer complexes.
Soluble HLA molecules and their modalities are extremely stable at 4˚C for several years and stay structurally intact up to a temperature of 42˚C.
Profit from short turnaround times, including target peptide validation, integrity and performance qualification and Mass-Spec exchange confirmation.
Talk to an expert today
Want to know more about HLA Engineering and its Application?
If you would still like to know more about our services or technology to advance your scientific discoveries and research, or are looking for some help in defining your project needs, reach out to our experts by visiting the Contact Us page.
Expert advice, whenever you need it!
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