Streamline your cell and gene therapy analytics

Streamline your cell and gene therapy analytics

Throughout the preclinical development and cell and gene therapy commercialization, analytical assays are essential to produce safe and potent drug products. Despite their importance, conventional analytical assays often consist of complex manual handling steps. Therefore, it can be unclear whether variations measured come from the drug product or if they have been unwittingly introduced by the operator or technology used. 
We have compiled a complete workflow solution for precise, easy-to-handle, and fast cell and gene therapy analytical assays to overcome these challenges in early discovery, cell therapy quality control during manufacturing, and patient monitoring research. Our solutions are applicable for different kinds of cell and gene therapies spanning a variety of cell type–specific applications, including but not limited to CAR T/NK, tumor-infiltrating leukocytes (TILs), regulatory T cells (Tregs), and induced pluripotent stem cells (iPSCs).

Browse through the chapters below and discover how standardization and automation enable a smooth transition from early discovery to clinical readiness or choose your cell type–specific application.

 

Early discovery and preclinical research of new therapies

The unprecedented efficacy of cell and gene therapies has encouraged the search for new targets and therapeutic approaches. This demanding task requires a versatile set of tools. To find relevant therapeutic targets samples must be prepared with care and analyzed with a high-throughput screening method. Newly designed drug products need to be characterized in detail and monitored within the tissue. Understanding the critical quality attributes required for treatment efficacy defines the clinical success of a new drug and establishing robust assays as early as possible is key for a seamless transition from research into the clinic. 

Gentle automated tissue dissociation

Conventional tissue dissociation methods can cause surface marker loss and decrease the viability of the cell suspension, which adversely affects sample analysis. Cells from solid tumors are especially difficult to preserve. 

Start smart with our innovative solutions for your tissue preparation and storage. We offer instruments, consumables, reagents, and support material for the automated preparation of up to eight tissue samples in parallel. By perfectly harmonizing enzymatic digestion and mechanical disruption, our gentleMACS™ Technology preserves epitope integrity and helps you standardize your tissue preparations to obtain reproducible data. 

Marker screening

Deep phenotyping of cancer cell diversity and the tumor microenvironment has been hampered by the lack of technologies that allow for multiplexed microscopy assays. 

The innovative MACSima™ Imaging Platform, together with the REAscreen™ MAX Kits, overcomes the limitations of the fluorescence spectrum utilizing a cyclic staining method and enables the visualization of >100 markers on a single tissue section. In addition, using our MACS Marker Screen, 378 pre-selected markers can be analyzed in a fast and easy manner by flow cytometry. 

Reliable cell analysis

Reliable immunophenotyping assays are essential to transfer a new cell and gene therapy from bench to bedside. Flow cytometry is the ideal tool for fast multiparametric surface marker analysis. Nevertheless, consistency in flow cytometric assays is restricted by lot-to-lot variations and non-specific binding of conventional hybridoma antibodies. 

Our unique REAfinity™ Recombinant Antibodies make reproducible and background-free cell analysis possible! Optimized and pre-tested antibody panels (Miltenyi Biotec Tested Panels: MBTPs) for a variety of CGT applications accelerate your panel design at no cost. Conjugation-optimized recombinant antibodies create opportunities for high-parameter analysis, such as mass cytometry or CITE-Seq. 

Conveniently reduce pipetting steps with our broad collection of ready-to-use flow cytometry kits and antibody cocktails for the phenotyping of critically relevant cell types for cellular therapies. Miltenyi Biotec offers immuno-oncology kits to characterize specific types of immune cells, for example, immune cell subsets in human blood or cell populations which have been identified to play great roles in the tumor microenvironment and may further lead to poor response of immunotherapy like MDSCs. In addition, several stem cell kits are available for fast and standardized characterization and quantification of cultured human MSCs or CD34+/CD133+ hematopoietic progenitor cells.

Application note
Background-free analysis of mouse TILs

Discover how REAfinity Recombinant Antibodies support your flow cytometric analysis by eliminating non-specific background signal.

Immunogenicity testing

Our reagents assist you in standardized preclinical assessment of immunogenicity against a variety of biological modalities, including protein therapeutics (e.g. monoclonal antibodies), cell therapeutics (e.g. genetically engineered cells or delivery systems), and gene therapeutics (e.g. virally delivered genes, oncolytic viruses, or CRISPR gene therapy). 

The analysis of immunogenicity is often focused on testing anti-drug antibody (ADA) formation. However, immunogenicity is caused by an antigen-specific T cell–dependent immune response. For this reason, early risk mitigation based on the assessment of, for example, therapeutic-specific T cell responses can increase the efficacy of your drug during the clinical stage. To ensure a robust analysis of these often rare antigen-specific T cells, we have developed MicroBeads and cell enrichment kits that allow efficient isolation and reliable analysis of viable antigen-specific T cells based on specific activation markers or cytokine expression. In addition, the rapid antigen-specific T cell enrichment (Rapid ARTE) protocol smoothly combines magnetic cell enrichment and multiparameter flow analysis for direct ex vivo detection and high-resolution characterization of rare antigen-specific T cells.

Imaging cells within 3D tumors

Commonly used techniques of classic histology and 2D microscopy are unable to meet the need for profound spatial analysis of large biological samples. However, high-quality 3D imaging of entire primary tumors, patient-derived xenografts, or large-scale organoid cultures, for example, to visualize the penetration of engineered immune cells into solid cancers, is a tall order. Our complete workflow, from antibodies specifically validated for 3D-immunofluorescence and deep tissue labeling for use with the MACS Clearing Kit, to the fully automated UltraMicroscope Blaze™ Light Sheet Imaging System, provides everything you need to visualize cells within the complex and three-dimensional tumor microenvironment.

Static 3D images of large samples give you a good overview – from subcellular details to whole organs or tumors. If you want to unravel dynamic processes like cell migration or cell-cell interactions, it can be useful to add time as a fourth dimension. Multiphoton microscopy shows what is happening in vivo in real time. Watch cells at work deep in living tissue using our TriM Scope™ Multiphoton Microscopes.

 

GMP manufacturing: in-process and quality control 

Streamlined and standardized cell therapy quality control is critical to decrease time to market and ensure delivery of a safe product. This entails reducing the number of work steps, therefore, donating hands-free time to the staff and avoiding human error. Cell manufacturing analytics need to reliably verify release criteria (e. g. identity, purity, safety, and potency), meet regulatory requirements, and support accelerated and safe product delivery.

Precise and robust cell characterization

Multiparametric flow cytometry is the ideal tool for in-process and quality control (IPC/QC) during manufacturing. Unfortunately, most flow-based assays require operator excellence to achieve reliable results.

Expert-tested recombinant antibody cocktails dried onto the bottom of a standard flow cytometry tube, called StainExpress™ Dry Antibody Cocktails, are a great way to foster consistency amongst different manufacturing sites and increase productivity – independently of operator experience. 

Accurate and flexible identification of CAR expression can be achieved by adding one of our ready-to-use CAR detection reagents optimized for background-free flow cytometry into the panel. In combination with a MACSQuant® Analyzer 16 flow cytometer full workflow automation and harmonization of multiple instruments with Smart Gain Technology becomes possible! 

Ready-made and customized application-specific software packages called Express Modes based on algorithms fed with real data offer robust gating decisions even in samples with donor variability. The MACSQuantify™ 21 CFR Part 11 option perfectly supports your GxP workflow by enabling FDA-compliant data submission. 

Does your manufacturing process require analytics tailored to your needs? Choose equipment that fits your specific needs with the help of our Custom Antibody Design Service.

Video
Standardizing flow cytometry assays in cell manufacturing

Standardizing flow cytometric assays in cell manufacturing faces significant challenges from variability along the workflow. A complete solution to this problem must address three main sources of disagreement: variability across flow cytometer instruments, operators, and reagent lots . If your flow cytometry partner can’t fix these issues, demand better! Ready to standardize your flow cytometry assays?

Cell functionality assays

A major challenge during quality control is the functional profiling of cells. Nevertheless, reliable and reproducible analysis of such parameters is crucial to predict treatment efficacy.

We offer apoptosis and cell viability dyes to perform antigen-specific in vitro killing assays and ensure cellular fitness of your product before infusion, suitable for high-throughput and automated flow cytometry. 

Flow-compatible multiplex assays help you determine the secretion of up to 12 effector cytokines and cytotoxic agents in a single sample, e.g, in co-culture supernatants of CAR T cells and their respective target cells. For the in vitro functional characterization of Tregs, we have developed the Treg Suppression Inspector. 

Does your heart beat for stem cell–based therapies? Choose our StemMACS™ HSC-CFU Assay Kit for a standardized flow cytometric readout to evaluate the differentiation potential of your HSCs. The StemMACS Trilineage Differentiation Kit is a fast and ready-to-use tool to analyze the differentiation potential of your ES and iPS cell lines whilst a variety of StemMACS Media can be used for MSC quality control. 

Article
CGTI Expert Roundtable – Overcoming challenges in analytical assays for cellular therapies

Jara Joedicke1, Eric Alonzo2, and Marsilius Mues3

1Senior scientist, Max Delbrück Center Berlin; 2Strategy Lead, Oncology, Analytical Development & CMC Lead, bbT369 Program, bluebird bio; 3R&D Group Leader at Miltenyi Biotec

Cell & Gene Therapy Insights 2021; 7(8), 1195–1206

Safety assessment

Working with cell-based products transduced with lentiviral vectors? Despite the great opportunities created by this technology, it also comes with potential safety risks. Hence, regulatory authorities demand certain analytical assays. 

Lentiviral vector copy numbers (VCN) can be determined precisely with the new MACS COPYcheck Kit and residual replication-competent lentiviruses (RCL) can rapidly be detected with the MACS LENTIcheck Kit (only compatible with lentiviral vectors provided by Lentigen and Miltenyi Biotec). 

 

Patient monitoring research during treatment

Cell and gene therapies do not end with infusion or implantation of the ready-made product. Longitudinal monitoring research is key to estimating both treatment success and severe side effects. Importantly, these assays need to be transferable to allow multicentric and inter-trial comparisons.

Cell expansion and persistence

With increasing time periods, it becomes more and more difficult to detect residual target cells in the blood or tissue. We have the perfect tools to overcome these hurdles. 

Explore colorful new possibilities with the MACSQuant Analyzer 16, a fully automated flow cytometer, and rely on consistent performance from lot to lot with our high-quality REAfinity Recombinant Antibodies. Our REAfinity Recombinant Technology, which is part of our antibodies and CAR detection reagents, is designed to eliminate background signals and increase sensitivity. In addition, serial sorting of rare cell populations, realized by the gentle MACSQuant® Tyto® Cell Sorter, is highly advantageous in the analysis of very small subpopulations, particularly in single-cell analyses.

Webinar
Follow-up of patients undergoing CAR T cell therapy: tools for immune monitoring

CAR T cell therapy has emerged as an important therapeutic option for hematological malignancies. A sensitive follow-up of patients undergoing CD19-targeted CAR T cell therapy is important to analyze, e.g., the in vivo expansion and persistence of these cells in the context of the clinical response of the patient. Furthermore, downstream analysis of CAR T cells isolated from the patients’ blood could help identify factors that distinguish responders from non-responders. In this webinar Prof. Boris Fehse from the University Clinic Hamburg-Eppendorf and Dr. Thomas Stübig from the University Clinic Kiel will give insights into their work.

In-depth analysis

Reliable assays for the analysis of critically low cell frequencies are hard to find. With our unique and gentle MicroBeads and Isolation Kits, specifically designed for the separation and enrichment of CD19 CAR+ cells, different immune cell types, and stem cell populations, you can overcome this hurdle and pave the way for reliable downstream genomic analysis. 

Immune responses to protein and peptide drugs can influence the course of treatment. Our rapid antigen-specific T cell enrichment (Rapid ARTE) protocol, utilizing our CD154 MicroBead Kits, allows you to enrich and analyze rare antigen-specific T cells which can cause adverse immunogenicity. Moreover, cytokine production determines toxicity and side effects, such as cytokine release syndrome (CRS). Shine a light on cytokine release during treatment with our cytokine detection assays for large-scale analysis and ultimate sensitivity (with or without enrichment). 

 

Cell type–specific applications

Choose one of the following cell types to find more detailed information for the analysis of your cell and gene therapy of interest