Nonspecific Adsorption Research Articles

Application of cellulose to chromatographic media: Cellulose dissolution, and media fabrication and derivatization

As the cornerstone of chromatographic technology, the development of high-performance chromatographic media is a crucial means to enhance the purification efficiency of biological macromolecules. Cellulose is a popular biological separation medium due to its abundant hydroxyl group on the surface, easy modification and, weak non-specific adsorption. In this paper, the development of cellulosic solvent systems, typical preparation methods of cellulosic chromatographic media, and the enhancement of chromatographic properties of cellulosic chromatographic media by polymeric ligand grafting strategies and their mechanism of action are reviewed. Ultimately, based on the current research status, a promising outlook for the preparation of high-performance cellulose-based chromatographic media was presented.

Ultra-sensitive and label-free detection of Escherichia coli O157:H7 using graphene-based field effect transistor modified with heat-denatured casein

Sensitive detection of Escherichia coli O157:H7 (E. coli O157:H7) in suspected water samples is critical for containing the water-borne outbreak, which may cause acute diarrhea and intestinal bleeding. Here, we reported an improved graphene-based field effect transistor modified with heat-denatured casein (GFET) for sensitively detecting E. coli O157:H7 in real water samples. The heat-denatured casein coated on the graphene surface was served as a probe linker to bind with the specific antibody of E. coli O157:H7, and as a blocker to avoid the interference signal arising from the non-specific adsorption between the graphene surface and impurities in real water samples. The GFET biosensor provided a wide concentration range of 1–107 CFU·mL−1 for cultured E. coli O157:H7 with a detection limit of 1 CFU·mL−1. This developed GFET biosensor can also discriminate E. coli O157:H7 from other interfering substances with a high selectivity. In addition, correlation analysis of the E. coli O157:H7 in real samples using the GFET biosensor were proved the high agreement with enzyme-linked immunosorbent assay. The mass-produced GFET biosensor can achieve ultra-sensitive and label-free detection for the E. coli O157:H7 in water sample, promising prospective environmental monitoring application in the future.

A flash signal amplification approach for ultrasensitive and rapid detection of single nucleotide polymorphisms in tuberculosis

In recent years, the demand for rapid, sensitive, and simple methods for diagnosing deoxyribonucleic acid (DNA) has grown due to the increase in the variation of infectious diseases. This work aimed to develop a flash signal amplification method coupled with electrochemical detection for polymerase chain reaction (PCR)-free tuberculosis (TB) molecular diagnosis. We exploited the slightly miscible properties of butanol and water to instantly concentrate a capture probe DNA, a single-stranded mismatch DNA, and gold nanoparticles (AuNPs) to a small volume to reduce the diffusion and reaction time in the solution. In addition, the electrochemical signal was enhanced once two strands of DNA were hybridized and bound to the surface of the gold nanoparticle at an ultra-high density. To eliminate non-specific adsorption and identify mismatched DNA, the self-assembled monolayers (SAMs) and Muts proteins were sequentially modified on the working electrode. This sensitive and specific approach can detect as low as attomolar levels of DNA targets (18 aM) and is successfully applied to detecting tuberculosis-associated single nucleotide polymorphisms (SNPs) in synovial fluid. More importantly, as this biosensing strategy can amplify the signal in only a few seconds, it possesses a great potential for point-of-care and molecular diagnosis applications.

Dopamine-Based Copolymer Bottlebrushes for Functional Adhesives: Synthesis, Characterization, and Applications in Surface Engineering of Antifouling Polyethylene.

Nonpolar materials like polyolefins are notoriously challenging substrates for surface modification. However, this challenge is not observed in nature. Barnacle shells and mussels, for example, utilize catechol-based chemistry to fasten themselves onto all kinds of materials, such as boat hulls or plastic waste. Here, a design is proposed, synthesized, and demonstrated for a class of catechol-containing copolymers (terpolymers) for surface functionalization of polyolefins. Dopamine methacrylamide (DOMA), a catechol-containing monomer, is incorporated into a polymer chain together with methyl methacrylate (MMA) and 2-(2-bromoisobutyryloxy)ethyl methacrylate (BIEM). DOMA serves as adhesion points, BIEM provides functional sites for subsequent "grafting from" reactions, and MMA provides the possibility for concentration and conformation adjustment. First, the adhesive capabilities of DOMA are demonstrated by varying its content in the copolymer. Then, terpolymers are spin-coated on model Si substrates. Subsequently, the atom transfer initiator (ATRP) initiating group is used to graft a poly(methyl methacrylate) (PMMA) layer from the copolymers, with 40% DOMA content providing a coherent PMMA film. To demonstrate functionalization on a polyolefin substrate, the copolymer is spin-coated on high-density polyethylene (HDPE) substrates. A POEGMA layer is grafted from the ATRP initiator sites on the terpolymer chain on the HDPE films to provide antifouling characteristics. Static contact angle values and Fourier transform infrared (FTIR) spectra confirm the presence of POEGMA on the HDPE substrate. Finally, the anticipated antifouling functionality of grafted POEGMA is demonstrated by observing the inhibition of nonspecific adsorption of the fluorescein-modified bovine serum albumin (BSA) protein. The poly(oligoethylene glycol methacrylate) POEGMA layers grafted on 30% DOMA-containing copolymers on HDPE show optimal antifouling performance exhibiting a 95% reduction of BSA fluorescence compared to nonfunctionalized and surface-fouled polyethylene. These results demonstrate the successful utilization of catechol-based materials for functionalizing polyolefin surfaces.

A brushed magnetic microbeads carrier with host-guest structures for enhancing bio-detection performance in beads-based flow cytometry

Here we report a brushed magnetic microbeads (BMBs) carrier with host-guest structures for enhancing bio-detection performance in beads-based flow cytometry (FCM). The micro-sized MBs are employed as host spheres and the nano-sized spherical polyelectrolyte brushes (SPBs) are utilized as guest particles to achieve the covalent assembly of SPBs on MBs (that is BMBs). Our proposed BMBs carrier exhibits a remarkable signal to noise improvement of approximately 8–9 times in beads-based immunoassay on FCM compared to conventional MBs via preserving capture antibodies’ activity and reducing detection antibodies’ nonspecific adsorption. This study demonstrates the promising potential of our developed BMBs carrier for beads-based flow cytometry.

A tumor targeted antifouling upconversion nanoplatform for fluorescence imaging and precise photodynamic therapy triggered by NIR laser

Photodynamic therapy (PDT) has been considered as a promising treatment in the biomedical field because of low toxicity to normal tissues and minor trauma area. However, the PDT effect of materials is greatly affected by many factors, such as nonspecific adsorption and poor light penetration, etc. In this work, an intelligent nano platform has been constructed based on upconversion nanoparticles (UCNPs) loaded with a large amount of photosensitizers Ce6, which could specifically light up tumor tissues and stimulate the production of reactive oxygen species (ROS) under 980 nm near-infrared (NIR) irradiation, exhibiting a conspicuous imaging and therapeutic effect of PDT treatment for deep tumors. An excellent anti-fouling performance in complex biological substrate was obtained upon the judicious introduction of anti-fouling peptide, which also contributed to the improved PDT efficiency. In addition, the specificity of nanoplatform to malignant breast cancer cells was realized by modification of polypeptide targeting for HER2. This anti-fouling nanoplatform provided an original paradigm for the development of fluorescence imaging and PDT for deep tumor tissue with high targeting and therapeutic efficacy, promising to be used in the early therapy of malignant breast cancer specifically.

Dual Stealth Functional Immuno-magnetic Nanoparticles for High-Performance Isolation of Circulating Tumor Cells.

As a biomarker of hepatocellular carcinoma (HCC) biopsy, circulating tumor cells (CTCs) are often used in the diagnosis of cancer and treatment guidance. For CTCs detection, immuno-magnetic nanoparticles (IMNs) are one of the most commonly used platforms. However, the nonspecific adsorption of proteins and non-tumor cells weakens the performance of IMNs to capture CTCs. In this work, we developed an IMNs platform which was constructed by a biomimetic protein corona precoating and a polyethylene glycol (PEG) spacer to form the PEG and corona-coated IMNs (IP-CMNs). Due to the dual stealth effect of protein corona precoating and PEG spacer, the nonspecific protein adsorption and cell binding of P-CMNs could reduce by ∼5.5- and ∼5.4-fold, respectively, compared with those of unmodified particles. Furthermore, the PEG spacer could not only reduce the interaction between IP-CMNs and leukocytes but also enhance the capture performance toward tumor cells. By using artificial blood samples, the capture efficiency of IP-CMNs toward rare CTCs was found to be 88.3%, while it was 70.5% by using commercial IMNs. Finally, CTCs were successfully isolated in all HCC patient blood samples (7/7) using IP-CMNs. These results provide insight into the use of the multifunctional nanoplatform as a useful tool for CTCs detection.

Aptamer-based carbohydrate antigen 125 sensor with molybdenum disulfide functional hybrid materials

Epithelial ovarian cancer is a malignant tumor of the female reproductive system with insidious symptoms, aggressiveness, risk of metastasis, and high mortality. Carbohydrate antigen 125 (CA125), a standard biomarker for screening epithelial ovarian cancer, can be applied to track cancer progression and treatment response. Here, we constructed an aptamer-based electrochemical biosensor to achieve sensitive detection of CA125. Molybdenum disulfide (MoS2) was used as the stable layered substrate, combined with the irregular branched structure of gold nanoflowers (AuNFs) to provide the sensing interface with a large specific surface area by one-step electrodeposition AuNFs@MoS2. The simplified electrode modification step increased the stability of the electrode while ensuring excellent electrochemical performance and providing many sulfhydryl binding sites. Then, AuNFs@MoS2/CA125 aptamer/MCH sensor was designed for CA125 detection. Based on AuNFs@MoS2 electrode, CA125 aptamer with sulfhydryl as the sensitive layer was fixed on the electrode by gold sulfur bonds. 6-Mercapto-1-hexanol (MCH) was used to block the electrode and reduce the non-specific adsorption. Finally, DPV analysis was applied for CA125 detection with the range of 0.0001 U/mL to 500 U/mL. Our designed aptamer sensor showed reasonable specificity, reproducibility, and stability. Clinical sample testing also proved the consistency of our sensor with the gold standard in negative/positive judgment. This work demonstrated a novel strategy for integrating nanostructures and biocompatibility to build advanced cancer biomarker sensors with promising applications.

Photoelectrochemical aptasensor based on PTCDA/TiO2 efficient sensitization effect for sensitive protein analysis

Herein, a photoelectrochemical (PEC) aptasensor for sensitive detection of C-reactive protein (CRP) was designed based on the efficient sensitization effect of 3,4,9,10-Perylene tetracarboxylic dianhydride (PTCDA) towards titanium dioxide (TiO2). Initially, TiO2 and PTCDA were continuously incubated on electrode surface to produce a strong PEC signal owing to the PTCDA/TiO2 sensitization structure. Subsequently, electrodeposition of gold nanoparticles (dep Au) was performed to connect with the incubated CRP aptamer (S1) via AuN bonds. After that, 1-hexanethiol (HT) was added on the surface of the modified electrode to block the non-specific adsorption sites. Eventually, with the addition of CRP, the PEC signal decreased significantly because the CRP-S1 complexes generated by the specific recognition of CRP and S1 resulted in high steric hindrance effect and inhibited electron transfer. Under optimal conditions, the proposed PEC aptasensor was applied successfully for sensitive CRP detection with a wide linear range from 10 pg/mL to 10 μg/mL and a low detection limit of 3.33 pg/mL. Moreover, the designed PEC aptasensor based on PTCDA/TiO2 sensitized structure might be a versatile tool that provides a powerful alternative to traditional proteins, ions and RNAs detection methods by changing the corresponding recognition sequences.

A 3D sliding-strip microfluidic device for the simultaneous determination of multiple target analytes

A simple paper-based microfluidic device was fabricated to simultaneously detect multiple targets. Microfluidic paper-based analytical devices (μPAD) comprise a single-layer moving sliding PAD (SPAD) to control the flow channel switch together with a folding origami PAD (OPAD) to test the target analytes. The facile assembly without any splicing materials avoids cross-contamination and non-specific adsorption of joining materials that may be caused by multi-target detection. The concentration of Fe(III), Ni(II), Cr(VI), and nitrite in standard solutions and actual aqueous solutions was successfully determined using the designed μPAD. The μPAD was able to achieve LOD of 3.3 mg/L, 1.3 mg/L, 0.35 mg/L, 0.28 mg/L for Fe (III), Ni (II), Cr (VI), and nitrite, respectively. The designed SOPAD exhibits improved stability, with a deviation of less than 7% compared to conventional analytical methods (ICP-OES and UV). Our work demonstrates that this 3D PAD holds great promise and a wide scope in environmental monitoring, biochemical analysis, food testing and other testing industries.

Multifunctional Fluorinated Lubricant Infused Poly(4-hydroxybutyrate) (P4HB) Membranes for Full Thickness Abdominal Wall Defect Repair.

Abdominal wall defect caused by surgical trauma, congenital rupture, or tumor resection may result in hernia formation or even death. Tension-free abdominal wall defect repair by using patches is the gold standard to solve such problems. However, adhesions following patch implantation remain one of the most challenging issues in surgical practice. The development of new kinds of barriers is key to addressing peritoneal adhesions and repairing abdominal wall defects. It's already well recognized that ideal barrier materials need to have good resistance to nonspecific protein adsorption, cell adhesion, and bacterial colonization for preventing the initial development of adhesion. Herein, electrospun poly(4-hydroxybutyrate) (P4HB) membranes infused with perfluorocarbon oil were used as physical barriers. The oil-infused P4HB membranes can greatly prevent protein attachment and reduce blood cell adhesion in vitro. We further showed that the perfluorocarbon oil-infused P4HB membranes can reduce bacterial colonization. The in vivo study revealed that perfluoro-(decahydronaphthalene) infused P4HB membranes can significantly prevent peritoneal adhesions in the classic abdominal wall defects model and accelerate defect repair, as evidenced by gross examination and histological evaluation. This work provides a safe fluorinated lubricant-impregnated P4HB physical barrier to inhibit the formation of postoperative peritoneal adhesions and efficiently repair soft-tissue defects. This article is protected by copyright. All rights reserved.

Preparation of chitin/MXene/poly(L-arginine) composite aerogel spheres for specific adsorption of bilirubin

Currently, hemoperfusion is clinically the most rapid and effective treatment for removing toxins from the blood. The core of hemoperfusion is the sorbent inside the hemoperfusion device. Due to the complex composition of the blood, adsorbents tend to adsorb substances such as proteins in the blood (non-specific adsorption) while adsorbing toxins. Hyperbilirubinemia is caused by excessive levels of bilirubin in the human blood, causing irreversible damage to the patient's brain and nervous system, and even leading to death. High adsorption and high biocompatibility adsorbents with specific bilirubin adsorption are urgently needed to treat hyperbilirubinemia. Herein, poly(L-arginine) (PLA) which can specifically adsorb bilirubin, was introduced into chitin/MXene (Ch/MX) composite aerogel spheres. Ch/MX/PLA prepared by supercritical CO2 technology had higher mechanical properties than Ch/MX and can withstand 50,000 times its own weight. The in vitro simulated hemoperfusion test showed that the adsorption capacity of Ch/MX/PLA was as high as 596.31 mg/g, which was 15.38 % higher than that of Ch/MX. Binary and ternary competitive adsorption tests showed that Ch/MX/PLA also had good adsorption capacity in the presence of a variety of interfering molecules. In addition, hemolysis rate testing and CCK-8 testing confirmed that Ch/MX/PLA had better biocompatibility and hemocompatibility. Ch/MX/PLA can meet the required properties of clinical hemoperfusion sorbents and has the ability to produce mass production. It has good application potential in the clinical treatment of hyperbilirubinemia.

Column Watch: Methods for the Passivation of HPLC Instruments and Columns

Metal-analyte interaction is a type of nonspecific adsorption (NSA) that occurs mainly between acidic analytes and active adsorptive sites present in the metal surfaces of the fluidic path of any chromatographic system. NSA represents a chromatographic challenge since it may lead to poor peak shape, low recovery, and poor performance from the high performance liquid chromatography (HPLC) system. As a result, many solutions that circumvent and mitigate this type of secondary interaction have been published or commercialized in the last few years. Such solutions encompass swapping stainless steel components for non-reactive polymers, “biocompatible” metals or alloys, the addition of additives into the mobile phase, pretreating the system with large amounts of sample or strong acids, and the use of coated stainless steel materials. This article explores these solutions in more detail, particularly the recent adaptation and benefits of coated metal surfaces in LC.

Anti-Fouling Strategies of Electrochemical Sensors for Tumor Markers.

The early detection and prognosis of cancers require sensitive and accurate detection methods; with developments in medicine, electrochemical biosensors have been developed that can meet these clinical needs. However, the composition of biological samples represented by serum is complex; when substances undergo non-specific adsorption to an electrode and cause fouling, the sensitivity and accuracy of the electrochemical sensor are affected. In order to reduce the effects of fouling on electrochemical sensors, a variety of anti-fouling materials and methods have been developed, and enormous progress has been made over the past few decades. Herein, the recent advances in anti-fouling materials and strategies for using electrochemical sensors for tumor markers are reviewed; we focus on new anti-fouling methods that separate the immunorecognition and signal readout platforms.

Do not let electrode fouling be the enemy of bioanalysis

Electrochemical biosensors are one of the best choices for the sensing of biomarkers due to their high performance, low cost, miniaturization, and wide applicability. However, like for any sensing process, electrode fouling affects severely the analytical performance of the sensor, such as sensitivity, detection limit, reproducibility, and overall reliability. Fouling arises from nonspecific adsorption of different components present in the sensing medium and in particular in complex biofluids such as full blood. The complex composition of blood where biomarkers are present at extremely low concentrations compared to the rest of the fluid composition makes electrochemical biosensing challenging. Direct biomarker analysis within full blood samples remains however central for the future development in electrochemical-based diagnostics. Herein, we aim to provide short discussion of past and more recent strategies and concepts employed to diminish background noise due to surface fouling and overcome current hurdles for the implementation and commercialisation of electrochemical-based biosensors for medical diagnostics of protein biomarkers in a point of care format.

Engineering Biomimetic Biosensor Using Dual-Targeting Multivalent Aptamer Regulated 3D DNA Walker Enables High-Performance Detection of Heterogeneous Circulating Tumor Cells.

The phenotypic heterogeneity of circulating tumor cells (CTCs) and the nonspecific adsorption of background cells impede the effective and sensitive detection of rare CTCs. Although leukocyte membrane coating approach has a good antileukocyte adhesion ability and holds great promise for addressing the challenge of capture purity, its limited specificity and sensitivity prevent its use in the detection of heterogeneous CTCs. To overcome these obstacles, a biomimetic biosensor that integrated dual-targeting multivalent aptamer/walker duplex functionalized biomimetic magnetic beads and an enzyme-powered DNA walker signal amplification strategy is designed. As compared to conventional leukocyte membrane coating, the biomimetic biosensor achieves efficient and high purity enrichment of heterogeneous CTCs with different epithelial cell adhesion molecule (EpCAM) expression while minimizing the interference of leukocytes. Meanwhile, the capture of target cells can trigger the release of walker strands to activate an enzyme-powered DNA walker, resulting in cascade signal amplification and the ultrasensitive and accurate detection of rare heterogeneous CTCs. Importantly, the captured CTCs remained viable and can be recultured in vitro with success. Overall, this work provides a new perspective for the efficient detection of heterogeneous CTCs by biomimetic membrane coating and paves the way for early cancer diagnosis.

Localized surface plasmon resonance biosensor chip surface modification and signal amplifications toward rapid and sensitive detection of COVID-19 infections

We developed a multi-pronged approach to enhance the detection sensitivity of localized surface plasmon resonance (LSPR) sensor chips to detect SARS-CoV-2. To this end, poly(amidoamine) dendrimers were immobilized onto the surface of LSPR sensor chips to serve as templates to further conjugate aptamers specific for SARS-CoV-2. The immobilized dendrimers were shown to reduce surface nonspecific adsorptions and increase capturing ligand density on the sensor chips, thereby improving detection sensitivity. To characterize the detection sensitivity of the surface-modified sensor chips, SARS-CoV-2 spike protein receptor-binding domain was detected using LSPR sensor chips with different surface modifications. The results showed that the dendrimer-aptamer modified LSPR sensor chip exhibited a limit of detection (LOD) of 21.9 pM, a sensitivity that was 9 times and 152 times more sensitive than the traditional aptamer- or antibody-based LSPR sensor chips, respectively. In addition, detection sensitivity was further improved by combining rolling circle amplification product and gold nanoparticles to further amplify the detection signals by increasing both the target mass and plasmonic coupling effects. Using pseudo SARS-CoV-2 viral particles as detection targets, we demonstrated that this combined signal intensification approach further enhanced the detection sensitivity by 10 folds with a remarkable LOD of 148 vp/mL, making it one of the most sensitive SARS-CoV-2 detection assays reported to date. These results highlight the potential of a novel LSPR-based detection platform for sensitive and rapid detection of COVID-19 infections, as well as other viral infections and point-of-care applications.

Molecular-electromechanical system for unamplified detection of trace analytes in biofluids.

Biological research and diagnostic applications normally require analysis of trace analytes in biofluids. Although considerable advancements have been made in developing precise molecular assays, the trade-off between sensitivity and ability to resist non-specific adsorption remains a challenge. Here, we describe the implementation of a testing platform based on a molecular-electromechanical system (MolEMS) immobilized on graphene field-effect transistors. A MolEMS is a self-assembled DNA nanostructure, containing a stiff tetrahedral base and a flexible single-stranded DNA cantilever. Electromechanical actuation of the cantilever modulates sensing events close to the transistor channel, improving signal-transduction efficiency, while the stiff base prevents non-specific adsorption of background molecules present in biofluids. A MolEMS realizes unamplified detection of proteins, ions, small molecules and nucleic acids within minutes and has a limit of detection of several copies in 100 μl of testing solution, offering an assay methodology with wide-ranging applications. In this protocol, we provide step-by-step procedures for MolEMS design and assemblage, sensor manufacture and operation of a MolEMS in several applications. We also describe adaptations to construct a portable detection platform. It takes ~18 h to construct the device and ~4 min to finish the testing from sample addition to result.

Affinity-based electrochemical biosensor with antifouling properties for detection of lysophosphatidic acid, a promising early-stage ovarian cancer biomarker

Electrochemical techniques are considered to be highly sensitive, capable of fast response and can be easily miniaturized, properties which can aid with regard to the fabrication of compact point-of-care medical devices; however, the main challenge in developing such a tool is overcoming a ubiquitous, problematic phenomenon known as non-specific adsorption (NSA). NSA is due to the fouling of non-target molecules in the blood on the recognition surface of the device. To overcome NSA, we have developed an affinity-based electrochemical biosensor using medical-grade stainless steel electrodes and following a unique and novel strategy using silane-based interfacial chemistry to detect lysophosphatidic acid (LPA), a highly promising biomarker, which was found to be elevated in 90 % of stage I OC patients and gradually increases as the disease progresses to later stages. The biorecognition surface was developed using the affinity-based gelsolin-actin system, which was previously investigated by our group to detect LPA using fluorescence spectroscopy. We demonstrate the capability of this label-free biosensor to detect LPA in goat serum with a detection limit of 0.7 µM as a proof-of-concept for the early diagnosis of ovarian cancer.

Enhanced Long-Term Antifouling Ability and Enrichment Effect of a Vertically Ordered Mesoporous Silica Film via Covalent Linkage of Chondroitin Sulfate for In Situ Detection of Cu2+ in Real Environmental Samples

The design and construction of antifouling sensing interfaces have been widely concerned due to the matrix effect of the complex environment and nonspecific adsorption at the electrode interface. However, it is challenging to ensure the long-term stability of an antifouling sensing interface during real sample detection. Herein, an interface construction strategy based on covalent bonds is proposed for striking enhancement of the long-term antifouling ability of a sensing interface based on a vertically ordered mesoporous silica film (VMSF) and a biobased antifouling material─chondroitin sulfate (CS). The covalent linkage of VMSF with CS (named VMSF-g-CS/GCE) could strengthen the long-term hydrophilicity and biocompatibility of VMSF/GCE. As expected, the VMSF-g-CS/GCE exhibited excellent long-term antifouling performances, which is a promising candidate sensing platform to be applied to long-term monitoring. Alternatively, the VMSF-g-CS/GCE exhibits a better enrichment effect to Cu2+ than VMSF/GCE and VMSF-CS/GCE (without covalent bonds). Therefore, VMSF-g-CS/GCE exhibited a current response toward Cu2+ within the scope of 0.1–0.8 μM, with a limit of detection (LOD) of 20.58 nM. The proposed design strategy offers an idea for the precise regulation of antifouling sensing surfaces, and the resulting high-performance sensing platform is attractive for in situ detection of Cu2+.