A comparative study on EpCAM antibody immobilization on gold surfaces and microfluidic channels for the detection of circulating tumor cells

Çetin, Didem; Okan, Meltem; Bat, Erhan; Külah, Haluk

doi:10.1016/j.colsurfb.2020.110808

Cited by 20 publications

(10 citation statements)

References 42 publications

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“…In the second case, streptavidin and biotinylated antibodies were used. The results indicated that the presence of long aromatic chains containing alkanethiols promoted better cell-uptaking [16].…”

Section: Discussionmentioning

confidence: 97%

“…In a study by Cetin et al, two techniques of gold surface modification were presented to immobilize anti-EpCAM antibodies to detect CTCs. The authors used for modification of self-assembled monolayers (SAMs) [16], which reduce non-specific binding of molecules regard to pure metal [34]. The antibodies were bound in two ways -by covalent bonds and by a bioaffinity interaction.…”

Section: Discussionmentioning

confidence: 99%

“…In order to do this, the carrier must be covered with an intermediate layer, interacting with the antibodies by the formation of covalent bonds or through bioaffinity interaction. Such chemical or topographic modifications of solid phase carriers allow them to bind antibodies more efficiently and achieve higher yields of captured cells [16].…”

Section: Introductionmentioning

confidence: 99%

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Positive influence of aminosilanes on anti-EpCAM antibody immobilization on a glass surface

Kamińska

Buszka

Pietras

et al. 2021

Medical Journal of Cell Biology

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Immobilization of antibodies has a number of promising applications, including detection of biomolecules and cells. Well-oriented antibodies are required to bind them effectively. To eliminate the problem of random antibodies’ orientation, the surface of the device can be modified with silanes. This study aimed at elucidating if selected aminosilanes were able to bind antibodies in the appropriate orientation and thus retain their binding activity. Silanization of glass slides was performed using three amino-functional trialkoxysilanes – A, AE, and AEE. The immunofluorescent reaction was used to evaluate the potential of the silanized glass surface to bind anti-EpCAM antibodies. The affinity of selected anti-EpCAM HEA125 antibodies labeled with fluorochrome to tested silanized surfaces was evaluated by measuring the mean fluorescence intensity (MFI) in each analyzed area. The presented silanes effectively bound antibodies. Higher fluorescence intensity was noticed in the case of silane-coated glass slides in comparison to unmodified ones. The differences in the contact angles also confirmed this result. In the case of silane A, the fluorescence intensity reflected the amount of bound antibodies. However, there was no such a relation in the case of the silanes AE and AEE. Although our research gave promising results, the usefulness of selected silanes needs to be confirmed by further studies using cancer cells. Running title: Aminosilanes as enhancers of antibody immobilization

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Section: Discussionmentioning

confidence: 97%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Positive influence of aminosilanes on anti-EpCAM antibody immobilization on a glass surface

Kamińska

Buszka

Pietras

et al. 2021

Medical Journal of Cell Biology

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“…Results showed that the presence of the aromatic ring in the within alkanethiols allowed for intermolecular interactions and consequently increased cell capture events. Additionally, covalent bonding permitted high flow rates within the microchannels, with negligible numbers of detached cells [ 74 ]. Furthermore, Chan et al investigated the efficiency of PMMA microfluidic channels coated with EpCAM antibodies-immobilized polyoxazoline polymer through covalent bonds for the photodynamic identification of EJ138, HT1376, HT1197, and RT4 human bladder cancer cells treated with protoporphyrin IX.…”

Section: Diagnostic Applicationsmentioning

confidence: 99%

Biosensors-on-Chip: An Up-to-Date Review

et al. 2020

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Generally, biosensors are designed to translate physical, chemical, or biological events into measurable signals, thus offering qualitative and/or quantitative information regarding the target analytes. While the biosensor field has received considerable scientific interest, integrating this technology with microfluidics could further bring significant improvements in terms of sensitivity and specificity, resolution, automation, throughput, reproducibility, reliability, and accuracy. In this manner, biosensors-on-chip (BoC) could represent the bridging gap between diagnostics in central laboratories and diagnostics at the patient bedside, bringing substantial advancements in point-of-care (PoC) diagnostic applications. In this context, the aim of this manuscript is to provide an up-to-date overview of BoC system development and their most recent application towards the diagnosis of cancer, infectious diseases, and neurodegenerative disorders.

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“…Over the years ELISA has significantly improved in its specificity and sensitivity, reaching pM concentrations [9,10], and since their first diagnostic application in 1971, the range of platforms in which they are used has largely expanded. Antibody applications range from attaching them to fluorescent [11,12], gold, or magnetic beads [13][14][15][16][17], to many activated surfaces, gold, glass, carbon [18][19][20][21].…”

Section: Introductionmentioning

confidence: 99%

Enhancing Antibodies’ Binding Capacity through Oriented Functionalization of Plasmonic Surfaces

et al. 2021

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Protein A has long been used in different research fields due to its ability to specifically recognize immunoglobulins (Ig). The protein derived from Staphylococcus aureus binds Ig through the Fc region of the antibody, showing its strongest binding in immunoglobulin G (IgG), making it the most used protein in its purification and detection. The research presented here integrates, for the first time, protein A to a silicon surface patterned with gold nanoparticles for the oriented binding of IgG. The signal detection is conveyed through a metal enhanced fluorescence (MEF) system. Orienting immunoglobulins allows the exposition of the fragment antigen-binding (Fab) region for the binding to its antigen, substantially increasing the binding capacity per antibody immobilized. Antibodies orientation is of crucial importance in many diagnostics devices, particularly when either component is in limited quantities.

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A comparative study on EpCAM antibody immobilization on gold surfaces and microfluidic channels for the detection of circulating tumor cells

Cited by 20 publications

References 42 publications

Positive influence of aminosilanes on anti-EpCAM antibody immobilization on a glass surface

Positive influence of aminosilanes on anti-EpCAM antibody immobilization on a glass surface

Biosensors-on-Chip: An Up-to-Date Review

Enhancing Antibodies’ Binding Capacity through Oriented Functionalization of Plasmonic Surfaces

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