Recombinant antibodies for flow cytometry – Miltenyi Biotec

Advantages of REAfinity Recombinant Antibodies

Flow cytometry is in their genes

REAfinity Antibodies are recombinant antibodies that provide superior lot-to-lot consistency and purity compared to mouse or rat hybridoma-derived, monoclonal antibodies. They have been recombinantly engineered to produce highly specific antibodies that require no FcR blocking step. Additionally, they all have the same IgG1 isotype, requiring less isotype controls.

REAfinity Recombinant Antibody model

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Performance data of recombinant antibodies

Reproducible results through consistent antibody quality

Mass spectrometry analysis of purified antibodies shows that REAfinity Antibodies are defined products, while hybridoma generated antibodies can be a mixture.

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The manufacturing process of REAfinity Recombinant Antibodies ensures high performance consistency based on the following aspects:

They are derived from a defined DNA sequence, which encodes only one type of heavy and light chain, ensuring high antibody purity.
Production under biologically and chemically defined, standardized culture conditions in mammalian cells, resulting in high lot-to-lot consistency.
Analytical, biochemical, and cell-based testing of every manufactured lot.

Only one isotype control – save efforts when setting up experiments

REAfinity Antibodies reduce the complexity of panel design with only one isotype control required.

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All REAfinity Recombinant Antibody clones contain the same specifically mutated human IgG1 sequence as their constant region. This offers the possibility to work with only one type of isotype control, reducing the complexity of experiment planning and saving time.

Hybridoma-derived monoclonal antibodies on the other hand, are composed of antibody isotypes derived from different species and therefore require multiple isotype controls. This complicates not only the management of reagent inventory, but also panel design when setting up experiments.

Optimized for flow cytometry applications

Specific detection of human CD116⁺ cells with clone REA 211.

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REAfinity Recombinant Antibodies are designed with flow cytometry in mind. The following properties ensure their superior performance as compared to traditional hybridoma-derived monoclonal antibodies:

Greater purity
No background signal
Highly optimized and standardized fluorochrome conjugation process

No more background signal – analyze what matters

Staining with REAfinity Antibodies shows no background signal, even without FcR block.

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All REAfinity Recombinant Antibodies contain a specifically mutated human IgG1 Fc region that abolishes their binding to Fcγ receptors. This allows for background-free analysis and eliminates the need for additional blocking steps such as using a FcR blocking reagent.

“We have seen that there is a huge advantage in terms of the background signal.”

More expert opinions

Dr. Anis Larbi

Head of the flow cytometry platform at the A*STAR Institute
Singapore

Fluorophore options

Flexibility in flow cytometry with thousands of recombinant antibodies

REAfinity Recombinant Antibody clones are available for hundreds of specificities.They are conjugated to a variety of fluorochromes to address multicolor flow cytometry needs. Current options include traditional dyes, like FITC, PE, APC, aa well as Miltenyi Biotec´s proprietary Vio® and VioBright™ Dyes.

Emission spectra of Vio Dyes: VioBlue, VioBright 515, Vio 515, VioGreen, VioBright FITC, PE-Vio 615, PerCP-Vio 700, PE-Vio 770, APC-Vio 770

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Videos

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Video
Recombinant antibodies in flow cytometry

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Video
Cap’n T Cell explains REAfinity Recombinant Antibodies

Let Cap’n T Cell take you on a ride through the wonderful world of REAfinity Recombinant Antibodies. In this video, he will explain the benefits of using REAfinity Antibodies for flow cytometry experiments and will show you the features that make these antibodies so unique.

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Webinar
Recombinant antibodies for improved flow cytometry

Join Dr. Kalpana Singh and her webinar on the advantages of recombinantly generated REAfinity antibodies that can enhance the reproducibility of your results and improve flow cytometry analysis

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Documentary
Antibody validation challenges

The documentary highlights the concerns raised by key stakeholders to understand if this recent awareness in reproducibility is truly a crisis or an opportunity to lay down some guidelines that will strengthen our faith in scientific data and outcomes going forward.

Resources

PDF downloads

REAfinity Recombinant Antibodies (brochure)
Recombinant antibodies for improved standardization in flow cytometry (scientific poster)

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