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Pro5® MHC Class I Pentamers

ProT2® MHC Class II Tetramers

ProM2® MHC Class II Monomers

CD1d Tetramers

thinkpeptides® Custom Peptide Synthesis

Antibodies

ProM1™ MHC Class I Monomers

ProMix™ Peptide Pools

ProPresent® Antigen Presentation Assay

ProSentium® Antigen Presentation Data

ProMap® T Cell Proliferation Assays

ProScern® DC-T Cell Proliferation Assays

ProImmune REVEAL & ProVE

ProStorm® Cytokine Release Assay

MutaMap® Mutational Activity Map

ProArray Ultra® B Cell Linear Epitope Mapping

ProArray Ultra® Ligand Binding Assays

T cell ELISPOT Assays

B cell ELISPOT Assay Service

Immunocytometry Assay Service

Blood Processing Service

typeHLA: HLA Typing Service

Special Assay Service

Immunogenicity of Biotherapeutics

Immunotoxicology Testing

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Updates

January 2019

Mastering Immunity Europe 2019 speakers announced and registration open

April 2015

ProImmune Introduces ProSentium™ a Unique Peptide Sequence Database to Investigate T Cell Immune System

New service and extensive data collection covers vast range of antigens with high disease relevance

Oxford,UK, 14 April 2015 – ProImmune, a leader in services for understanding and managing adaptive immune responses, announced today the introduction of a new antigen database service called ProSentium™, based on in vitro assay results using sequencing mass spectrometry. ProSentium data can show the precise peptide sequences from proteins of interest visible to the body’s own T cell immune system. The data collection covers many high value therapeutic targets and areas including the majority of licensed replacement factor proteins, including clotting factors, replacement enzymes, spanning development areas including oncology, cardiovascular, CNS, immunology, gastroenterology and many orphan diseases.

Under an exclusive or non-exclusive license, the bespoke database service investigates and reports peptide hits against the ProSentium database for a customer’s specific protein or protein family of interest. Results are reported as overlapping nested peptide sets and provide statistically relevant antigen sequence data that can be used to further inform drug development decisions.

Nikolai Schwabe, CEO of ProImmune commented: “ProSentium supports pivotal decision making in drug design and development with the physical evidence required for game changing approaches that target or interact with the immune system. Through our service offering, we are delighted that we can offer this ground-breaking resource to researchers so they can better understand the body’s immune response and bring new therapies to patients. ”

T cells play a critical role in all immune responses, including in fighting infections, cancer and in autoimmunity and in unwanted drug reactions. To understand T cell responses, it is crucial to understand the specific peptide antigens that the immune system recognizes and are presented on the cell surface of both normal and diseased cells. ProSentium data can be readily analyzed and reported, and is fully compatible with ProImmune’s in vitro assay services which can be combined to help investigators gain a more complete understanding of relevant immune responses at a molecular level.

For more information on ProSentium please visit: www.proimmune.com, email enquiries@proimmune.com , or call +1 888 505 7765 in the United States or +44 870 042 7279 inother countries.

ProImmune Introduces ProT2^™MHC Class II Tetramer Reagents to Study Antigen-Specific CD4+ T CellImmune Responses

Oxford, UK, 14 January 2016

ProImmune, a leader in services for understanding and managing adaptive immune responses, announced today the introduction of ProT2^™, a range of Human MHC Class II tetramer reagents for tetramer analyses in the study of antigen-specific CD4+ T cell immune responses. ProImmune’s offering represents the broadest choice of ready to order catalog and custom items in this field and will enable research into CD4+ T cell responses in a variety of disease areas including cancer, infectious diseases, allergy, autoimmunity and transplantation. Another key area for this product range is the detailed study of T cell help in unwanted immune responses to biologics.

CD4+ T cell responses are at the heart of steering the adaptive immune system directing both cytotoxic T cell responses and antibody responses mediated by B cells. ProT2™ MHC Class II Tetramers allow the direct detection and separation of single antigen-specific CD4+T cells accurately by flow cytometry.

Nikolai Schwabe, CEO of ProImmune commented: “By introducing a broad and growing ProT2™ Tetramer product range we intend to give MHC Class II tetramer analysis the emphasis it deserves in the study of CD4+ T cell responses which play such a pivotal role in adaptive immunity across so many disease and treatment modalities.”

ProT2™ Class II MHC Monomer reagents are supported by ProImmune’s expert customer service and application support, helping new users in particular to establish and optimize robust protocols in their own laboratories.

This addition to ProImmune’s class II reagent offering further complements an already extensive range of services for evaluating CD4+ T cell responses, including ProImmune REVEAL® MHC peptide binding assays, CD4+ T cell and DC T cell proliferation assays and standard immune monitoring services through ELISPOT and flow cytometry based assays.

For more information on ProT2™ reagents and their availability, or to place an order, please visit: www.proimmune.com ,email enquiries@proimmune.com , or call + 1 888 505 7765 in the United States or + 44 870 042 7279 in other countries.

-ENDS-

Notes to Editors:

About ProImmune - www.proimmune.com

ProImmune’s mission is to be a partner of choice for understanding and managing adaptive immune responses. It does this by offering unique solutions for preclinical and clinical immunology research, including a comprehensive and integrated antigen characterization and immunogenicity testing platform, and products and services for tracking antigen-specific immune responses with state-of-the-art ELISpot and flow cytometry techniques. ProImmune is committed to helping researcher’s achieve success through product innovation, responsive service and focused application support, saving time and money, and reducing risk.

For more information, please contact

At ProImmune:

Jeremy Fry,

Tel: +44 (0) 870 042 7279

Email: enquiries@proimmune.com

Media enquiries:

Sarah Jeffery

Zyme Communications Ltd

Tel: +44 (0) 7771 730 919

Email: sarah.jeffery@zymecommunications.com

September 2018

MHC Pentamers used by team at GSK to study mRNA vaccines that can protect against several Flu strains

Pro5® Class I MHC Pentamers

used by team at GSK to study mRNA vaccines that can protect against several Flu strains Magini D. et al., PLOS One (2017) Self-Amplifying mRNA Vaccines Expressing Multiple Conserved Influenza Antigens Confer Protection against Homologous and Heterosubtypic Viral Challenge http://dx.doi.org/10.1371/journal.pone.0161193

Figure: NP-specific CD8⁺ T-cell responses in lungs after influenza challenge. BALB/c mice were immunized i.m. twice, 8 weeks apart, with PBS, 0.1 μg of self-amplifying mRNA (SAM®) vectors SAM(NP), SAM(M1), SAM(M1-NP), or with 0.2 μg of SAM(NP)+SAM(M1). Four weeks after the second immunization, mice were infected with PR8 virus. NP-specific CD8 T cells recruited in the lungs after the infection were characterized by flow cytometry. (a) Numbers of NP-specific CD8⁺ T cells determined using Pro5® MHC pentamers. Data are from individual mice (depicted as dots), while solid lanes indicate the mean±SD. (b) Cumulative frequency of Ag-specific, cytokine-secreting CD8⁺ T cells, indicated as absolute number per lung. The color code represents the different combinations of cytokine produced by the respective cells after in vitro stimulation with medium (m), NP_147-155 peptide (NP), or M1 peptide pool (M1), as indicated. (c) Absolute number of NP-specific CD8⁺ T cells positive (black bar) or not (grey bar) for CD107a. Data derived from two independent and merged experiments. Statistical analyses were performed using the Mann-Whitney U test. *p<0.05; **p<0.01 compared to the PBS-treated group. http://dx.doi.org/10.1371/journal.pone.0161193.g006 Request a Quote Abbreviated abstract: Current hemagglutinin (HA)-based seasonal influenza vaccines induce vaccine strain-specific neutralizing antibodies that usually fail to protect against mismatched circulating viruses. Inclusion of conserved proteins such as the nucleoprotein (NP) and the matrix protein 1 (M1) can increase effectiveness by eliciting cross-reactive T-cells. However, efficient priming of T-cell responses requires the right delivery system. Here we show novel self-amplifying mRNA (SAM^®) vectors expressing influenza NP (SAM(NP)), M1 (SAM(M1)), and NP and M1 (SAM(M1-NP)) delivered with lipid nanoparticles (LNP) induce robust polyfunctional CD4 T helper 1 cells, while NP-containing SAM also induced cytotoxic CD8 T cells. Robust expansions of central memory (T_CM) and effector memory (T_EM) CD4 and CD8 T cells were also measured. An enhanced recruitment of NP-specific cytotoxic CD8 T cells was observed in the lungs of SAM(NP)-immunized mice after influenza infection that paralleled with reduced lung viral titers and pathology, and increased survival after homologous and heterosubtypic influenza challenge. We show for the first time that the co-administration of RNA (SAM(M1-NP)) and protein (monovalent inactivated influenza vaccine (MIIV)) was feasible, induced simultaneously NP-, M1- and HA-specific T cells and HA-specific neutralizing antibodies, and enhanced MIIV efficacy against a heterologous challenge. Pro5® MHC Pentamer

Pro5® MHC Class I Pentamers are the market leading product for detecting antigen-specific T cells, you can count on consistent performance backed up by unrivalled customer service. To ensure uniformity and consistency, each Pentamer is purified four times before being subjected to rigorous quality control acceptance testing.

The choice of leading researchers
Hundreds of catalog items
Custom made for new specificities
Uniform bright staining
Excellent long-term stability
More than 1200 scientific publications
Better uniform avidity than tetramers
Unrivalled application support

Request a Quote

USA & Canada, Toll Free: +1 888 505 7765 All other countries: +44 (0)870 042 7279 Distributors Email: enquiries@proimmune.com More on Pro5® Class I MHC Pentamers Customer Publications Subscribe to Email Newsletters

January 2018

ProImmune launches MutaMap™ Mutational Activity Map Service

MutaMap™ Mutational Activity Map

high throughput service for mutational activity maps of therapeutic proteins

A new key tool for developability engineering.

When it’s time to make final engineering decisions for your antibody or protein, MutaMap™ can help evaluate which individual point mutations to pursue. MutaMap™ is an in vitro assay system that helps explore the effect of substituting each amino acid in at each position in a protein sequence one by one with all 19 possible substitutions and find out the effect on protein activity. For example a sequence stretch of 100 amino acids will result in up to 2000 mutants to explore. The approach of MutaMap™ is simple. Each position of a protein of interest is mutated by site directed mutagenesis, expressed and tested for its affinity/activity. MutaMap™ does not use any surrogate measurement for affinity or activity. Cell free in vitro translation of proteins is combined with solution titration assays to measure affinity/activity. Both methods are optimized for high throughput processing of samples while still allowing for accurate measurements of affinity/activity. The technology is particularly suitable for investing the ligand binding interactions of high affinity monoclonal antibodies, down into the high femtomolar range where other approaches such as SPR struggle to deliver high throughput results.

Watch our MutaMap™ Podcast Mutations investigated in CDR1 and CDR3 of Avastin® heavy chain variable domain

Figure 1: Shown above is an example of a mutagenesis heatmap generated for CDR1 and CDR3 from Avastin® heavy chain variable fragment. For binding pair interactions MutaMap™ uses high throughput solution equilibrium titration (SET) immunoassays to determine the binding affinity for each construct tested.

Figure 2: SET results for measuring the affinity of Avastin® scFv and Lucentis® scFv. The resulting titration curve is regressed according to the relevant mass action binding laws. The robustness of the SET approach ensures that this high throughput assay works well for affinities in the single digit picomolar range, as demonstrated by the tight confidence intervals. What does MutaMap™ show you? MutaMap™ delivers a heat map for your protein (see Figure 1 above) that shows you which point mutations lead to an increase, decrease, or no change in affinity (or other activity) or non-function of the protein when interacting with one or more of its binding partners. Effectively you can learn which mutations, one by one, are likely to be permissible or favourable in your protein in terms of the key property of binding to a ligand. MutaMap™ therefore allows you to make informed protein engineering decisions for a range of key developability objectives which include:

Improving activity/affinity by cherry picking mutations, including in the CDRs of mAbs
De-immunization while maintaining or even improving protein activity
Altering cross reactivity, e.g. for improved cross-species reactivity
Improved humanization or provision of other engineered features
Stability/Manufacturability and other key developability improvements, achievable through deliberate point mutations
Prolonging half-life
Developing unique new composition of matter IP

Focus on position S105 in CDR-H3 of Avastin® heavy chain variable domain

Figure 3: What is clear is that the MutaMap™ heatmap shows permissible mutations, especially in CDR-H3 in cases that are not normally considered conservative, e.g. in position S105. This opens up choices for re-designing the molecule that would not normally be available based on computational assessments.

Figure 4: Position S105 in Avastin® CDR-H3 is mutated to T in in Lucentis® MutaMap™ reconfirms that this mutation is indeed beneficial for improving binding. It also shows that a number of other mutations are available to match or improve the affinity of the construct over the Avastin® wild type.

How does MutaMap™ compare to molecular evolution technologies?

Molecular evolution techniques such as phage display and other phenotype-genotype coupled randomization techniques are most commonly used in the affinity maturation process for monoclonal antibodies and other binding scaffolds. The advantage of these technologies is that they help explore a very large sequence space of combined mutations.

There comes a point however when final decisions have to be made on the implementation of a protein sequence where individual point mutations may be considered in an antibody or therapeutic protein to meet a variety of design objectives. Randomized molecular evolution is not appropriate for this step. Exploring individual point mutations is nothing new, but it has been difficult to carry this step out in very high throughput way, especially where the objective is to clone and express every mutant and then measure its affinity/activity with reasonable accuracy. This is what MutaMap™ can achieve.

Where does MutaMap™ fit in as part a project for e.g. generating a candidate monoclonal antibody for clinical development?

Figure 5:Example work flow for pre-clinial protein engineering of therapeutic monoclonal antibody; individual projects may differ.

How long does a MutaMap™ project take?

Our objective is to complete medium size projects of exploring 500-2000 mutations in approximately 8-12 weeks from receiving the customer’s protein sequence. Larger projects will take slightly longer, depending on complexity.

What is the stepwise process for carrying out MutaMap™?

First we will discuss with you the sequence space you want to explore. This may be the known paratope of a protein or the CDR and flanking regions of a monoclonal antibody.
To save time and cost you may not want to explore substituting amino acids into the sequence that are considered highly non-conservative or prone to degradation when exposed. We will also discuss what you know about the binding interaction of your protein with its target, whether you have working immunoassays for this interaction and whether the wild type protein and isolated ligand is available to work with in an immunoassay system. Depending on the nature of the interaction and the binding partners we will ensure that the base assay of binding wild type protein to target works well.
Once these details are agreed and the project is initiated we will run the mutagenesis agreed for each position, express the protein at small scale and carry out the equilibrium binding titration affinity measurement in high throughput.

What will you get?

A final technical report delivered via our secure webserver showing you the affinity or activity determination for the wild type and each mutant with confidence interval. This will be presented in various formats for ease of interpretation, including a standard heatmap.

For customers that want to carry out protein antigenicity studies in parallel, these can be carried out in approximately the same timeframe as the MutaMap™. This means that within a period of approximately 8-12 weeks we will have determined experimentally both the putative T cell epitopes and the MutaMap™ of permitted mutations in your protein sequence. This information can allow you to proceed with much better informed decisions on how to address immunogenicity related issues for your program while addressing simultaneously other developability related design decisions for your sequence.

More News

05 Aug 2019 To 09 Aug 2019

Immuno-Oncology Summit, Boston

21 Mar 2019 To 21 Mar 2019

Mastering Immunity Europe 2019, Oxford

08 Apr 2019 To 12 Apr 2019

PEGS Summit 2019, Boston

09 May 2019 To 13 May 2019

AAI 2019, San Diego

18 Mar 2019 To 22 Mar 2019

Immuno-Oncology Summit Europe 2019, London

17 Jun 2019 To 20 Jun 2019

18th Annual World Preclinical Congress, Boston

21 Oct 2019 To 25 Oct 2019

Immunogenicity & Bioassay Summit, VA

09 Sep 2019 To 10 Sep 2019

Mastering Immunity 2019, Museum of Science, Boston

18 Nov 2019 To 22 Nov 2019

PEGS Europe 2019, Lisbon

25 Feb 2019 To 27 Feb 2019

European Immunogenicity Platform, Lisbon

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MutaMap™ Mutational Activity Map

ProStorm® Cytokine Release Assay

ProT2® MHC Class II Tetramers: Detecting Antigen-specific CD4+ T cells

ProScern®: DC-T Cell Proliferation Assay

ProMap®: Naïve T Cell Proliferation Assay

ProSentium™: The T cell Immuno-Epiproteome at your fingertips

ProImmune REVEAL and ProVE®

Dr Jeremy Fry provides a summary of ProImmune’s REVEAL and ProVE® service, the rapid epitope discovery system.

Pro5® MHC Pentamers

Dr Andrew Waller provides an overview of the Pro5 Pentamer technology and the benefits of using MHC pentamers to detect specific CD8 T Cell populations in your research.