Towards precision medicine: the role and potential of protein and peptide microarrays

Brambilla, Dario; Chiari, Marcella; Gori, Alessandro; Cretich, Marina

doi:10.1039/c9an01142k

Cited by 17 publications

(11 citation statements)

References 117 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…While common chemical linking approaches, targeting amine‐, carboxyl‐, or thiol‐ groups usually work effortless, they cannot control the specific orientation of AB in relation to the surface, as these groups are available in abundance on the AB. [ 55–57 ] Therefore, a fraction of the ABs will be oriented with their active site toward the surface, thus rendering them inactive for capture. Certain click‐chemistry approaches as well as making use of biochemical interactions (e.g., streptavidin‐biotin, protein A/G binding to ABs) can achieve an orientation‐controlled immobilization.…”

Section: Resultsmentioning

confidence: 99%

“…Certain click‐chemistry approaches as well as making use of biochemical interactions (e.g., streptavidin‐biotin, protein A/G binding to ABs) can achieve an orientation‐controlled immobilization. [ 56 ] However, these approaches are usually more elaborate, and, e.g., in the streptavidin‐biotin strategy, also the ABs need to be biotinylated site‐specifically, otherwise orientation is again random. All standard approaches have in common, that they bring the AB usually into very near vicinity to the surface with only a linker molecule in between.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Rapid Capture of Cancer Extracellular Vesicles by Lipid Patch Microarrays

et al. 2021

View full text Add to dashboard Cite

Extracellular vesicles (EVs) contain various bioactive molecules such as DNA, RNA, and proteins, and play a key role in the regulation of cancer progression. Furthermore, cancer‐associated EVs carry specific biomarkers and can be used in liquid biopsy for cancer detection. However, it is still technically challenging and time consuming to detect or isolate cancer‐associated EVs from complex biofluids (e.g., blood). Here, a novel EV‐capture strategy based on dip‐pen nanolithography generated microarrays of supported lipid membranes is presented. These arrays carry specific antibodies recognizing EV‐ and cancer‐specific surface biomarkers, enabling highly selective and efficient capture. Importantly, it is shown that the nucleic acid cargo of captured EVs is retained on the lipid array, providing the potential for downstream analysis. Finally, the feasibility of EV capture from patient sera is demonstrated. The demonstrated platform offers rapid capture, high specificity, and sensitivity, with only a small need in analyte volume and without additional purification steps. The platform is applied in context of cancer‐associated EVs, but it can easily be adapted to other diagnostic EV targets by use of corresponding antibodies.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Rapid Capture of Cancer Extracellular Vesicles by Lipid Patch Microarrays

et al. 2021

View full text Add to dashboard Cite

show abstract

“…First, although it is now straightforward and economical to chemically synthesize many peptides, it is commonly believed that insufficiently large numbers of ECSPs can be identified for a given antigen. Second, peptides are relatively more prone to NSIs, which has to date severely complicated or even preclude the development of diagnostic kits (Huang et al, 2015) (Brambilla et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

“…While there are many studies related to NSI (James et al, 2003) (Eisen and Chakraborty, 2010) (Brambilla et al, 2019), surprisingly, we are unaware of any research examining the reproducibility of Ab-Ag interactions. The reason for this lack of scrutiny may be due to the fact that the magnitude of any given mAb-probe interaction, either specific or non-specific, can be predicted with a thermodynamic equilibrium equation ([Ab-Ag] = [Ab][Ag]/K D ), which, as a model, also implies the intrinsically reproducible characteristics of the Ab-Ag interactions.…”

Section: Introductionmentioning

confidence: 99%

Previously unrecognized non-reproducible antibody-antigen interactions and their implications for diagnosis of viral infections including COVID-19

Pan

Yang

Deng

et al. 2021

Preprint

View full text Add to dashboard Cite

Antibody-antigen (Ab-Ag) interactions are canonically described by a model which exclusively accommodates non-interaction (0) or reproducible-interaction (RI) states, yet this model is inadequate to explain often-encountered non-reproducible signals. Here, by monitoring diverse experimental systems and confirmed COVID-19 clinical sera using a peptide microarray, we observed that non-specific interactions (NSI) comprise a substantial proportion of non-reproducible antibody-based results. This enabled our discovery and capacity to reliably identify non-reproducible Ab-Ag interactions (NRI), as well as our development of a powerful explanatory model ("0-RI-NRI-Hook four-state model") that is [mAb]-dependent, regardless of specificity, which ultimately shows that both NSI and NRI are not predictable yet certain-to-happen. In experiments using seven FDA-approved mAb drugs, we demonstrated the use of NSI counts in predicting epitope type. Beyond challenging the centrality of Ab-Ag interaction specificity data in serology and immunology, our discoveries also facilitated the rapid development of a serological test with uniquely informative COVID-19 diagnosis performance.

show abstract

“…In this scenario, peptide microarrays are appealing tools as they allow the simultaneous screening of hundreds to thousands different peptides immobilized in spatially ordered patterns of spots on solid supports 9 10 possibly leading to the development of synthetic diagnostic probes with improved specificity and of putative, new immunotherapeutic targets.…”

Section: Introductionmentioning

confidence: 99%

SARS-CoV-2 epitope mapping on microarrays highlights strong immune-response to N protein region

Musicò

Frigerio

Mussida

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

A workflow for SARS-CoV-2 epitope discovery on peptide microarrays is herein reported. The process started with a proteome-wide screening of immunoreactivity based on the use of a high-density microarray followed by a refinement and validation phase on a restricted panel of probes using microarrays with tailored peptide immobilization through a click-based strategy. Progressively larger, independent cohorts of Covid-19 positive sera were tested in the refinement processes, leading to the identification of immunodominant regions on SARS-CoV-2 Spike (S), Nucleocapsid (N) protein and Orf1ab polyprotein. A summary study testing 50 serum samples highlighted an epitope of the N protein (region 155-171) providing 92% sensitivity and 100% specificity of IgG detection in Covid-19 samples thus being a promising candidate for rapid implementation in serological tests.

show abstract

Towards precision medicine: the role and potential of protein and peptide microarrays

Abstract: Techniques to comprehensively analyze protein signatures are pivotal to unravel disease mechanisms, develop novel biomarkers and targeted therapies. In this frame, protein and peptide microarrays can play a major role in fuelling precision medicine.

Cited by 17 publications

References 117 publications

Rapid Capture of Cancer Extracellular Vesicles by Lipid Patch Microarrays

Rapid Capture of Cancer Extracellular Vesicles by Lipid Patch Microarrays

Previously unrecognized non-reproducible antibody-antigen interactions and their implications for diagnosis of viral infections including COVID-19

SARS-CoV-2 epitope mapping on microarrays highlights strong immune-response to N protein region

Contact Info

Product

Resources

About