Comprehensive in vitro characterization of PD-L1 small molecule inhibitors

Ganesan, Aravindhan; Ahmed, Marawan; Okoye, Isobel; Arutyunova, Elena; Babu, Dinesh; Turnbull, William L.; Kundu, Joydeb Kumar; Shields, Justin; Agopsowicz, Katharine Cheryl; Xu, Lai; Tabana, Yasser; Srivastava, Nutan; Zhang, Guangzhi; Moon, Tae Chul; Belovodskiy, Alexandr; Hena, Mostofa Abu; Kandadai, Appan Srinivas; Hosseini, Seyedeh Nargess; Hitt, Mary; Walker, John; Smylie, Michael; West, F. G.; Siraki, Arno G.; Lemieux, M. Joanne; Elahi, Shokrollah; Nieman, James A.; Tyrrell, D. Lorne; Houghton, Michael; Barakat, Khaled

doi:10.1038/s41598-019-48826-6

Cited by 98 publications

(100 citation statements)

References 54 publications

(62 reference statements)

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“…[24] In another recent publication, researchers describe a set of potent macrocyclic peptide and small-molecule PD-1/PD-L1 inhibitors, one of which induced cytokine production (IL-2 and IFN-γ) and T cell proliferation at levels comparable to pembrolizumab. [25] We suspect the unique binding properties observed for many of the antibodies studied here will translate to similarly unique biological activity at the cellular and potential therapeutic level.…”

Section: Plos Onementioning

confidence: 92%

Assessing the binding properties of the anti-PD-1 antibody landscape using label-free biosensors

et al. 2020

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Here we describe an industry-wide collaboration aimed at assessing the binding properties of a comprehensive panel of monoclonal antibodies (mAbs) against programmed cell death protein 1 (PD-1), an important checkpoint protein in cancer immunotherapy and validated therapeutic target, with well over thirty unique mAbs either in clinical development or market-approved in the United States, the European Union or China. The binding kinetics of the PD-1/mAb interactions were measured by surface plasmon resonance (SPR) using a Carterra LSA instrument and the results were compared to data collected on a Biacore 8K. The effect of chip type on the SPR-derived binding rate constants and affinities were explored and the results compared with solution affinities from Meso Scale Discovery (MSD) and Kinetic Exclusion Assay (KinExA) experiments. When using flat chip types, the LSA and 8K platforms yielded near-identical kinetic rate and affinity constants that matched solution phase values more closely than those produced on 3D-hydrogels. Of the anti-PD-1 mAbs tested, which included a portion of those known to be in clinical development or approved, the affinities spanned from single digit picomolar to nearly 425 nM, challenging the dynamic range of our methods. The LSA instrument was also used to perform epitope binning and ligand competition studies which revealed over ten unique competitive binding profiles within this group of mAbs.

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Section: Plos Onementioning

confidence: 92%

Assessing the binding properties of the anti-PD-1 antibody landscape using label-free biosensors

et al. 2020

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“…As per available literature, potential predictive biomarkers that can be used to select patients who may benefit from combined treatment using HER2-targeted and PD-1/PD-L1 axis based therapeutic agents are (1) HER2 amplification/overexpression, (2) PD-1/PD-L1 expression, (3) presence of a greater number of TILs and fewer Tregs, (4) higher TMB (tumor mutation burden), (5) PTEN expression, and (6) expression of CD5, CD74, CD96, and CD226, to name only few [38,86,121,[131][132][133][134][135][136][137][138]. However, it is still not clear which combination of clinicopathological factors are most reliable predictive biomarkers to implement effective treatment protocols using anti-HER2 and/or PD-1/PD-L1 pathways [39,139].…”

Section: Pd-1/pd-l1 and Her2 Crosstalk In Breast Cancermentioning

confidence: 99%

Crosstalk between HER2 and PD-1/PD-L1 in Breast Cancer: From Clinical Applications to Mathematical Models

Padmanabhan

Kheraldine

Meskin

et al. 2020

Cancers

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Breast cancer is one of the major causes of mortality in women worldwide. The most aggressive breast cancer subtypes are human epidermal growth factor receptor-positive (HER2+) and triple-negative breast cancers. Therapies targeting HER2 receptors have significantly improved HER2+ breast cancer patient outcomes. However, several recent studies have pointed out the deficiency of existing treatment protocols in combatting disease relapse and improving response rates to treatment. Overriding the inherent actions of the immune system to detect and annihilate cancer via the immune checkpoint pathways is one of the important hallmarks of cancer. Thus, restoration of these pathways by various means of immunomodulation has shown beneficial effects in the management of various types of cancers, including breast. We herein review the recent progress in the management of HER2+ breast cancer via HER2-targeted therapies, and its association with the programmed death receptor-1 (PD-1)/programmed death ligand-1 (PD-L1) axis. In order to link research in the areas of medicine and mathematics and point out specific opportunities for providing efficient theoretical analysis related to HER2+ breast cancer management, we also review mathematical models pertaining to the dynamics of HER2+ breast cancer and immune checkpoint inhibitors.

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“…In any case, one binding partner is fluorescent, while the other binding partner remains label-free. MST has also been used to characterize protein-small molecule interactions such as those involving aptamers (Entzian and Schubert, 2016) and PD-L1 inhibitors by BMS (Ganesan et al, 2019) or for high-throughput screening to identify hits (Rainard et al, 2018). Here, we describe a protocol using MST to determine the binding affinity of the PD-1/PD-L1 complex, which is involved in tumor escape processes, without purification of the target protein from cell lysates (Khavrutskii et al, 2013).…”

Section: Microscale Thermophoresis (Mst) Is a Biophysical Technique Tmentioning

confidence: 99%

Measurement of Protein-Protein Interactions through Microscale Thermophoresis (MST)

Magnez¹,

Thiroux²,

Tardy³

et al. 2020

BIO-PROTOCOL

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The binding interactions of PD-1 and PD-L1 have been studied by surface plasmon resonance (SPR) and isothermal titration calorimetry (ITC) over the past few years, but these investigations resulted in controversy regarding the values of the dissociation constant (Kd) (Freeman et al., 2000). MST is a powerful new method for the quantitative analysis of protein-protein interactions (PPIs) with low sample consumption. The technique is based on the movement of molecules along microscopic temperature gradients, and it detects changes in their hydration shell, charge or size. One binding partner is fluorescently labeled, while the other binding partner remains label-free. We used a protocol that allows the determination of the binding affinity by MST without purification of the target protein from the cell lysate. The application of this MST method to PD-1-eGFP and PD-L1-eGFP expressed in CHO-K1 cells allowed us, for the first time, to determine the affinity of the complex formed between PD-1 and its ligand PD-L1 during tumor escape. The protocol has a variety of potential applications for studying the interactions of proteins with small molecules.

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Comprehensive in vitro characterization of PD-L1 small molecule inhibitors

Cited by 98 publications

References 54 publications

Assessing the binding properties of the anti-PD-1 antibody landscape using label-free biosensors

Assessing the binding properties of the anti-PD-1 antibody landscape using label-free biosensors

Crosstalk between HER2 and PD-1/PD-L1 in Breast Cancer: From Clinical Applications to Mathematical Models

Measurement of Protein-Protein Interactions through Microscale Thermophoresis (MST)

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