A flow-proteometric platform for analyzing protein concentration (FAP): Proof of concept for quantification of PD-L1 protein in cells and tissues

Chou, Chao-Kai; Huang, Po-Jung; Tsou, Pei Hsiang; Wei, Yongkun; Lee, Heng-Huan; Wang, Ying-Nai; Liu, Yen-Liang; Shi, Colin; Yeh, Hsin Chih; Kameoka, Jun; Hung, Mien‐Chie

doi:10.1016/j.bios.2018.05.053

Cited by 7 publications

(3 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…An advantage of our assay is that it used ac-EHD fluid flow to reduce the non-specific binding and minimise the chances for false positive results. Recently, Chou et al reported a microfluidic platform with fluorescence readout which achieved highly sensitive detection of PD-L1 in the picomolar range(Chou et al 2018). However, fluorescence based assays are limited by high background signal due to photo-bleaching and difficult for multiplexing due to spectral overlapping.…”

mentioning

confidence: 99%

A SERS microfluidic platform for targeting multiple soluble immune checkpoints

Khondakar

Sina

Wuethrich

et al. 2019

Biosensors and Bioelectronics

View full text Add to dashboard Cite

Immune checkpoint blockade therapies are promising next generation immunotherapeutic treatments for cancer. Whilst sequential solid biopsies are an invaluable source of prognostic information, they are not feasible for monitoring therapeutic outcomes over time. Monitoring soluble immune checkpoint markers expression in body fluids could potentially be a better alternative. Current methods (e.g. ELISA) for detecting immunecheckpoint proteins mostly rely on the use of monoclonal antibodies which are expensive and time-consuming to manufacture and isolate. Herein, we report an integrated surface enhanced Raman scattering (SERS)-microfluidics device for the detection of immune checkpoint proteins which involves the use of i) nano yeast single chain variable fragment (scFv) as a promising alternative to monoclonal antibodies providing high stability at relative low-cost and simplicity for production, ii) graphene oxide functionalised surface to reduces the bio functionalization steps, thus avoiding the general paradigm of biotin-streptavidin chemistry and iii) a microfluidic platform enabling alternating current electrohydrodynamics (ac-EHD) induced nanomixing to enhance the target scFv binding and minimize the non-specific interactions. Specific and multiplex detection of immune checkpoint biomarkers is achieved by SERS based spectral encoding. Using this platform, we successfully demonstrated the detection of clinically relevant soluble immune checkpoints PD-1, PD-L1 and LAG-3 from as low as 100 fg/mL of analytes spiked in human serum.

show abstract

mentioning

confidence: 99%

A SERS microfluidic platform for targeting multiple soluble immune checkpoints

Khondakar

Sina

Wuethrich

et al. 2019

Biosensors and Bioelectronics

View full text Add to dashboard Cite

show abstract

“…PD-L1 is an ideal target because it activates PD-1 signaling and is preferentially overexpressed by tumor or tumor-associated microenvironment [63,64]. Immunohistochemical (IHC) staining of patient tumor specimens to examine PD-L1 protein is a simple and direct method to stratify patients for anti-PD-1/PD-L1 treatment [65,66].…”

Section: Pd-l1 N-linked Glycosylation In Clinical Diagnosismentioning

confidence: 99%

The impact of PD-L1 N-linked glycosylation on cancer therapy and clinical diagnosis

et al. 2020

Self Cite

View full text Add to dashboard Cite

N-linked glycosylation is one of the most abundant posttranslational modifications of membrane-bound proteins in eukaryotes and affects a number of biological activities, including protein biosynthesis, protein stability, intracellular trafficking, subcellular localization, and ligand-receptor interaction. Accumulating evidence indicates that cell membrane immune checkpoint proteins, such as programmed death-ligand 1 (PD-L1), are glycosylated with heavy Nlinked glycan moieties in human cancers. N-linked glycosylation of PD-L1 maintains its protein stability and interaction with its cognate receptor, programmed cell death protein 1 (PD-1), and this in turn promotes evasion of T-cell immunity. Studies have suggested targeting PD-L1 glycosylation as a therapeutic option by rational combination of cancer immunotherapies. Interestingly, structural hindrance by N-glycan on PD-L1 in fixed samples impedes its recognition by PD-L1 diagnostic antibodies. Notably, the removal of N-linked glycosylation enhances PD-L1 detection in a variety of bioassays and more accurately predicts the therapeutic efficacy of PD-1/PD-L1 inhibitors, suggesting an important clinical implication of PD-L1 N-linked glycosylation. A detailed understanding of the regulatory mechanisms, cellular functions, and diagnostic limits underlying PD-L1 N-linked glycosylation could shed new light on the clinical development of immune checkpoint inhibitors for cancer treatment and deepen our knowledge of biomarkers to identify patients who would benefit the most from immunotherapy. In this review, we highlight the effects of protein glycosylation on cancer immunotherapy using N-linked glycosylation of PD-L1 as an example. In addition, we consider the potential impacts of PD-L1 N-linked glycosylation on clinical diagnosis. The notion of utilizing the deglycosylated form of PD-L1 as a predictive biomarker to guide anti-PD-1/PD-L1 immunotherapy is also discussed.

show abstract

“…Still, many other proteins may influence the expression of cancer controlling genes and they may also need to be regulated to win the battle with cancer. The use of nanomaterials in cancer research, diagnostics, and therapy is rapidly growing and spans from the targeted drug delivery using nanocarriers [1][2][3][4], to imaging [5,6], including MRI enhancing [7], and to biosensing using various platforms, such as biosensors and sensor arrays [8], microfluidic devices [9,10], and assays [11][12][13], and others. Novel quantum devices and plasmonic effects have recently been discovered, including DNA-linked quantum dots [14,15] and fluorescence enhancement [16][17][18].…”

Section: Introductionmentioning

confidence: 99%

Gated Resonance Energy Transfer (gRET) Controlled by Programmed Death Protein Ligand 1

et al. 2020

View full text Add to dashboard Cite

The resonance energy transfer (RET) between an excited fluorescent probe molecule and a plasmonic nanoparticle (AuNP) has been investigated to evaluate the effect of protein molecules on the RET efficiency. We have found that the energy transfer to a functionalized AuNP can be modulated by a sub-monolayer film of programmed death-ligand 1 (PD-L1) protein. The interactions of PD-L1 with AuNP@Cit involve incorporation of the protein in AuNP shell and formation of a submonolayer adsorption film with voids enabling gated surface plasmon resonance energy transfer (SPRET). A model of the gated-RET system based on the protein size, estimated using Fisher–Polikarpov–Craievich density approximation, has been developed and can be utilized for other proteins, with minimum data requirement, as well. The value of the equilibrium constant KL determined for the Langmuir isotherm is high: KL = 1.27 × 108 M−1, enabling highly sensitive control of the gated-RET by PD-L1. Thus, with the gated-RET technique, one can determine PD-L1 within the dynamic range, extending from 1.2 to 50 nM. Moreover, we have found that the Gibbs free energy for PD-L1 binding to AuNP@Cit is −46.26 kJ/mol (−11.05 kcal/mol), indicating a strong adsorption with supramolecular interactions. The proposed gated-RET system, with the fluorescence intensity of the fluorophore probe molecule modulated by plasmonic quenching with AuNP and shielding of energy transfer by the adsorbed PD-L1 can be further developed for determination of PD-L1 in pharmaceutical formulations for immune checkpoint control in cancer therapy.

show abstract

A flow-proteometric platform for analyzing protein concentration (FAP): Proof of concept for quantification of PD-L1 protein in cells and tissues

Cited by 7 publications

References 33 publications

A SERS microfluidic platform for targeting multiple soluble immune checkpoints

A SERS microfluidic platform for targeting multiple soluble immune checkpoints

The impact of PD-L1 N-linked glycosylation on cancer therapy and clinical diagnosis

Gated Resonance Energy Transfer (gRET) Controlled by Programmed Death Protein Ligand 1

Contact Info

Product

Resources

About