How To Choose An Anti-PD-1 Antibody For Your Research

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PD-1 Introduction

Programmed death-1 (PD-1) is an inhibitory receptor expressed on T-cells that promotes apoptosis of inflammatory T-cells and inhibits apoptosis of anti-inflammatory regulatory T-cells, thereby promoting self-tolerance and preventing autoimmune diseases. To evade immune surveillance, tumor cells often exploit this system by over-expressing PD-L1, the ligand which binds to and activates PD-1. Anti-PD-1 monoclonal antibodies are used to block the binding of PD-L1 to PD-1 which allows the immune system to find and kill tumor cells.

Anti-PD-1 Antibody Reactivity

The first step in deciding which anti-PD-1 antibody fits your research needs is to identify which species reactivity is needed for your experimental model. Bio X Cell offers anti-mouse and anti-human PD-1 antibodies. Which to choose is based on your experimental model. If you are using mice expressing mouse PD-1 you want to choose an anti-mouse PD-1 antibody. If you are using human cells in culture or humanized mice expressing human PD-1 then you want to choose an anti-human PD-1 antibody.

Anti-mouse PD-1 Antibody Clones

Bio X Cell offers three distinct anti-mouse PD-1 antibody clones: RMP1-14, 29F.1A12, and J43. All three antibodies function through the same mechanism – they bind to PD-1 and sterically block the binding of PD-1 to PD-1 ligands, thereby blocking PD-1 signaling (as shown in the image above). These antibodies do not function through depleting PD-1 expressing cells.

All three antibodies are ideally suited for in vivo blocking of PD-1 signaling in mouse models and have extensive multi-year publication records backing this application. The differences between these antibodies lie in their PD-1 blocking capacities, their listed reported (published) applications, their host and isotype, and their origins.

With respect to their PD-1 blocking capacities, a recent publication titled “A Comparison of Murine PD-1 and PD-L1 Monoclonal Antibodies” compares the RMP1-14 and 29F.1A12 clones. The authors found that the 29F.1A12 clone has approximately 100-fold higher PD-1 blocking affinity than the RMP1-14 clone. The authors further conclude that 29F.1A12 most closely models human PD-1 therapeutic antibodies, which have a high affinity and blocking ability. Further details on this study can be found here.

Among these anti-PD-1 antibodies, the RMP1-14 clone has the most extensive publication record for in vivo blocking of PD-1 signaling. Importantly, however, the reported applications for the RMP1-14 antibody are limited to in vivo blocking of PD-1/PD-L signaling only.

Alternatively, the 29F.1A12 clone also has an extensive publication record for in vivo PD-1 blocking but can also be used for in vitro PD-1 neutralization, western blotting, immunohistochemistry, immunofluorescence, and flow cytometry. The J43 clone fits a similar description. This can be advantageous for researchers who wish to run in vitro or diagnostic experiments in parallel with in vivo experiments.

The next step in making the antibody selection that is best for your research is to search the publication record for each clone and find an example of published data that uses a similar experimental approach to your own. There may be one clone that is more often used for your specific mouse model and experimental set-up.

Bio X Cell provides a list of selected references for all our antibodies which can be found on each product page. Additional references can be found using search engines such as Google Scholar and PubMed. Searching for the clone name of the antibody in combination with experiment-specific search terms, for example, “RMP1-14 MC38 BALB/c”, is the best approach to finding applicable references. These references will help you select the antibody that best fits your research.

Detailed Comparison of anti-mouse PD-1 Antibody Clones

All of Bio X Cell’s InVivoPlus™ antibodies, including these anti-PD-1 clones, are rigorously tested to ensure that they are murine pathogen free, have an endotoxin level of <1EU/mg, and a protein aggregation level of <5%. These clones are also available in our cost effective InVivoMAb™ format. These alternatives are: RMP1-14 (BE0146), 29F.1A12 (BE0273), and J43 (BE0033-2). A detailed explanation of the differences between our InVivoPlus™ and InVivoMAb™ formats can be viewed here.

Recombinant anti-mouse PD-1 Antibody Clones

Bio X Cell also offers recombinant chimeric versions of our three anti-mouse PD-1 antibody clones. RecombiMAb™ antibodies are provided as chimeric antibodies with mouse constant regions instead of the typical rat or hamster constant regions. This ensures reduced immunogenicity in sensitive mouse models. Select RecombiMAb™ antibodies are also available in multiple Fc silenced versions. These Fc silenced versions have mutations in the Fc fragment of the antibody rendering them unable to bind to endogenous Fcγ receptors. Learn More About RecombiMAb™ Antibodies

Detailed Comparison of Recombinant anti-mouse PD-1 Antibody Clones

Anti-human PD-1 Antibody Clones

Bio X Cell also offers multiple anti-human PD-1 antibodies for use in humanized mouse models or immunoassays. These include clones J116 and J110 and Pembrolizumab and Nivolumab biosimilar antibodies.

Detailed Comparison of anti-human PD-1 Antibody Clones