Ultrasensitive ELISA using enzyme-loaded nanospherical brushes as labels.

Improving the detection sensitivity of enzyme-linked immunosorbent assay (ELISA) is of utmost importance for meeting the demand of early disease diagnosis. Herein we report an ultrasensitive ELISA system using horseradish peroxidase (HRP)-loaded nanospherical poly(acrylic acid) brushes (SPAABs) as labels. HRP was covalently immobilized in SPAABs with high capacity and activity via an efficient "chemical conjugation after electrostatic entrapment" (CCEE) process, thus endowing SPAABs with high amplification capability as labels. The periphery of SPAAB-HRP was further utilized to bind a layer of antibody with high density for efficient capture of analytes owing to the three-dimensional architecture of SPAABs. Using human chorionic gonadotrophin (hCG) as a model analyte, the SPAAB-amplified system drastically boosted the detection limit of ELISA to 0.012 mIU mL(-1), a 267-fold improvement as compared to conventional ELISA systems.

Covalent immobilization of proteins on 3D poly(acrylic acid) brushes: mechanism study and a more effective and controllable process.

Polymeric brushes have emerged as a novel 3D material platform that provides great amounts of binding sites for biomolecules. This paper investigates the covalent immobilization mechanism of protein by spherical poly(acrylic acid) brushes (SPAABs) in the widely adopted N-hydroxysuccinimide/N-(3-dimethyl-aminopropyl)-N'-ethylcarbodiimide hydrochloride (NHS/EDC) process. It was discovered that electrostatic interaction plays a crucial role in the covalent immobilization of protein. Due to the existence of 3D architecture and "Donnan effect", SPAABs exhibit quite different immobilization kinetics in comparison with conventional 2D materials. Under conditions favorable to electrostatic interaction, the effect of "electrostatic interaction induced covalent binding" was observed as a result of competitive immobilization by physical adsorption and chemical binding. On the basis of the mechanism study, a new "chemical conjugation after electrostatic entrapment" (CCEE) method was developed which set the chemical and physical immobilization process apart. A more effective and well-defined covalent immobilization was achieved. And the binding capacity can be tuned in a wide range (0-4.2 mg protein/mg SPAABs) with a high level of control.

Enhancement of enzymatic activity by magnetic spherical polyelectrolyte brushes: a potential recycling strategy for enzymes.

Interactions between amyloglucosidase and magnetic spherical polyelectrolyte brushes (MSPB) were studied by turbidimetric titration, which reveals reversible and tunable behaviors of pH-dependent enzyme-SPB binding. Quantitative thermodyanmic parameters including binding affinity and stoichiometry between enzyme and SPBs were further measured by isothermal titration calorimetry (ITC). A large amount of enzyme can be loaded in MSPB without loss of MSPB stability. We demonstrated that the enzymatic activity of amyloglucosidase bound in MSPB could be greatly enhanced (catalytic reaction rate, k(bound) = 1.36k(free)) compared to free enzyme acitivity in solution. This is tremendous improvement from other carrier systems that usually lead to a significant decrease of enzymatic activity. Both the high enzyme loading capacity and the enhancement of the catalytic activity probably arise from the Coulombic interactions between the enzyme and MSPB. These findings provide a practical strategy for enhancement of enzyme activity and enzyme recycling by MSPB.

The Effect of Expanded Vermiculite on the Fire Resistance of Waterborne Acrylic Coatings.

Due to their ability to prevent or slow the spread of fires, fire-retardant coatings are utilized as the main means of fire protection for steel structures, combining easy application and high economic efficiency. This study investigates the effects of the particle size and dosage of expanded vermiculite (EV) on the fire resistance and application performance of coatings. Ammonium polyphosphate, melamine, and pentaerythritol were used as intumescent fire-retardant systems, along with waterborne hydroxyl-modified acrylic resins as the film-forming substances. The properties of fire resistance coatings were tested via scanning electron microscope (SEM), X-ray diffractometry (XRD), thermogravimetric analysis (TGA), limiting oxygen index (LOI), and cone calorimetry. The excellent fire performance of the coatings with 3 wt.% 300-mesh EV was proven, exhibiting a relative expansion of 30.43 times. Moreover, the surface structure of the charcoal layer was dense. The total smoke production (TSP) and smoke concentration (TSR) were only 0.18 m2 and 0.25 m2/m2.

Binding between proteins and cationic spherical polyelectrolyte brushes: effect of pH, ionic strength, and stoichiometry.

Cationic spherical polyelectrolyte brushes (SPBs) were synthesized by photoemulsion polymerization, consisting of a polystyrene core with a diameter around 80 nm and a poly(2-aminoethylmethacrylate hydrochloride) (PAEMH) shell with a thickness from 10 to 50 nm densely grafted on the core surface. The binding of various proteins onto SPBs was observed by turbidimetric titration, dynamic light scattering (DLS), zeta potential, and isothermal titration calorimetry (ITC). The binding, aggregation, and releasing of proteins by SPB can be tuned by modulating pH. The pH regions of binding for bovine serum albumin (BSA), ß-lactoglobulin (BLG), and papain onto SPBs are markedly different and tunable by ionic strength and stoichiometry between protein and SPB. Binding energetics, affinity, and amount of various proteins onto cationic SPBs were determined by ITC. These findings lay the foundation for SPB applications in the protein purification and selective immobilization of different proteins, enzymes, and antibodies.

Temperature Responsive Diblock Polymer Brushes as Nanoreactors for Silver Nanoparticles Catalysis.

Metal nanoparticles are widely used in catalysis. Loading metal nanoparticles into polymer brushes has aroused wide attention, but regulation of catalytic performance still needs to be improved. The novel diblock polymer brushes, polystyrene@sodium polystyrene sulfonate-b-poly (N-isopropylacrylamide) (PSV@PSS-b-PNIPA) and PSV@PNIPA-b-PSS with reversed block sequence, were prepared by surface initiated photoiniferter-mediated polymerization (SI-PIMP) and used as nanoreactors to load silver nanoparticles (AgNPs). The block sequence caused the difference of conformation and further affected the catalytic performance. PSV@PNIPA-b-PSS@Ag was found to be able to control the amount of AgNPs exposed to external reactant of 4-nitrophenol at different temperatures to achieve regulation of the reaction rate due to the hydrogen bonds and further physical crosslinking between PNIPA and PSS.

Brain developmental and cortical connectivity changes in transgenic monkeys carrying the human-specific duplicated gene SRGAP2C.

Human-specific duplicated genes contributed to phenotypic innovations during the origin of our own species, such as an enlarged brain and highly developed cognitive abilities. While prior studies on transgenic mice carrying the human-specific SRGAP2C gene have shown enhanced brain connectivity, the relevance to humans remains unclear due to the significant evolutionary gap between humans and rodents. In this study, to investigate the phenotypic outcome and underlying genetic mechanism of SRGAP2C, we generated transgenic cynomolgus macaques (Macaca fascicularis) carrying the human-specific SRGAP2C gene. Longitudinal MRI imaging revealed delayed brain development with region-specific volume changes, accompanied by altered myelination levels in the temporal and occipital regions. On a cellular level, the transgenic monkeys exhibited increased deep-layer neurons during fetal neurogenesis and delayed synaptic maturation in adolescence. Moreover, transcriptome analysis detected neotenic expression in molecular pathways related to neuron ensheathment, synaptic connections, extracellular matrix and energy metabolism. Cognitively, the transgenic monkeys demonstrated improved motor planning and execution skills. Together, our findings provide new insights into the mechanisms by which the newly evolved gene shapes the unique development and circuitry of the human brain.

Chinese herbal medicine combined with cognitive-behavioural therapy for avoidant paruresis: a controlled trial.

Background: Avoidant paruresis is a common clinical condition in urology and psychosomatic medicine. However, it has limited treatment options that are safe and effective with few side effects. Aims: Our study aimed to investigate the effectiveness and safety of the Chinese herbal Yangxin Tongquan decoction combined with cognitive-behavioural therapy (CBT) for avoidant paruresis. Methods: Sixty-eight patients with avoidant paruresis were divided into a treatment group (33 patients) and a control group (35 patients). The control group was assigned 10 weeks of CBT and systematic desensitisation. In addition to CBT and systematic desensitisation, the treatment group was given the Chinese herbal Yangxin Tongquan decoction during the 10-week study. The Shy Bladder Syndrome Scale (SBS) and the Self-rating Anxiety Scale (SAS) were administered before and after treatment to measure any change. Results: The overall efficacy in the treatment group (n=30) was 80.0% vs 62.5% in the control group (n=33). Comparing pretreatment and post-treatment measures, both groups showed improvement in SBS scores and SAS scores (treatment group: t(SBS) =8.397, p(SBS) <0.001, t(SAS) =8.216, p(SAS)<0.001; control group: t(SBS) =6.802, p(SBS) <0.001, t(SAS)=5.171, p(SAS) <0.001). Moreover, both groups' SBS and SAS scores changed significantly over time (SBS scores: Ftime =118.299, p<0.001; SAS scores: Ftime =92.114, p<0.001). However, the treatment group performed better than the control group (SBS scores: Ftime*group =5.709, p=0.020; SAS scores: Ftime*group =7.235, p=0.009). Conclusions: The Chinese herbal Yangxin Tongquan decoction combined with cognitive-behavioural psychotherapy positively affects the treatment of avoidant paruresis without significant adverse effects.

Spherical polyelectrolyte brushes as a nanoreactor for synthesis of ultrafine magnetic nanoparticles.

Ultrafine magnetic nanoparticles (MNP, 1.4-5.8 nm) were generated within a nanoreactor of spherical polyelectrolyte brushes (SPB). SPB consist of a solid polystyrene (PS) core and densely grafted poly(acrylic acid) (PAA) chains. Due to strong chemical coordination between carboxyl groups in PAA and MNP surfaces, the obtained magnetic spherical polyelectrolyte brushes (MSPB) kept excellent stability and maintained pH sensitivity. The magnetic properties of MSPB were confirmed by a vibrating sample magnetometer (VSM). The size and the size dispersion of MNP can be modulated by varying adding sequences (conventional coprecipitation or reverse coprecipitation), or nanoreactor structure (with or without crosslinking). MNP content in MSPB could be adjusted by multicycle reactions. This new strategy makes it possible to synthesize ultrafine inorganic nanoparticles with tunable size in SPB.

Study on the Synthesis and Properties of Waterborne Polyurea Modified by Epoxy Resin.

The most notable features of polyurea are its fast reaction, energy-saving and high efficiency. In order to meet the needs of environmental protection, waterborne polyurea (WPUA) has become a research hotspot. However, the presence of hydrophilic groups in WPUA reduces its solvent resistance, heat resistance and mechanical properties. Therefore, it is necessary and valuable to develop a high-performance WPUA. In this study, epoxy-modified waterborne polyurea (WPUAE) emulsions were prepared using epoxy resin as a modifier. Fourier transform infrared spectroscopy (FT-IR) showed that E44 was successfully introduced into the molecular chain of WPUA. The WPUAE was tested for gel fraction, adhesion, contact angle, solvent resistance, tensile properties and thermal stability. The results showed that when the E44 content was 8 wt%, the performance of WPUAE was best, the adhesion of WPUAE coating film was 1.53 MPa, the gel fraction, water contact angle, water absorption, toluene absorption, tensile strength and decomposition temperature were 96.94%, 70.3°, 16.43%, 131.04%, 9.05 MPa and 365 °C, respectively. The results showed that epoxy resin as an emulsion modifier improved the comprehensive properties of WPUA.

Comparison of lncRNA Expression in the Uterus between Periods of Embryo Implantation and Labor in Mice.

Uterine function during pregnancy is regulated mainly by progesterone (P4) and estrogen (E2). Serum P4 levels are known to fluctuate significantly over the course of pregnancy, especially during embryo implantation and labor. In this study, pregnant mice at E0.5, E4.5, E15.5, and E18.5 (n = 3/E) were used for an RNA-Seq-based analysis of mRNA and lncRNA expression. In this analysis, 1971 differentially expressed (DE) mRNAs, 493 known DE lncRNAs, and 1041 novel DE lncRNAs were found between E0.5 and E4.5 at the embryo implantation stage, while 1149 DE mRNAs, 192 known DE lncRNAs, and 218 novel DE lncRNAs were found between E15.5 and E18.5 at the labor stage. The expression level of lncRNA-MMP11 was significantly downregulated by P4 treatment on MSM cells, while lncRNA-ANKRD37 was significantly upregulated. Notably, 117 DE mRNAs, 19 known DE lncRNAs, and 31 novel DE lncRNAs were commonly expressed between the two stages, indicating that these mRNAs and lncRNAs may be directly or indirectly regulated by P4.

Protein purification by polyelectrolyte coacervation: influence of protein charge anisotropy on selectivity.

The effect of polyelectrolyte binding affinity on selective coacervation of proteins with the cationic polyelectrolyte, poly(diallyldimethylammonium chloride) (PDADMAC), was investigated for bovine serum albumin/ß-lactoglobulin (BSA/BLG) and for the isoforms BLG-A/BLG-B. High-sensitivity turbidimetric titrations were used to define conditions of complex formation and coacervation (pH(c) and pH(ϕ), respectively) as a function of ionic strength. The resultant phase boundaries, essential for the choice of conditions for selective coacervation for the chosen protein pairs, are nonmonotonic with respect to ionic strength, for both pH(c) and pH(ϕ). These results are explained in the context of short-range attraction/long-range repulsion governing initial protein binding "on the wrong side of pI" and also subsequent phase separation due to charge neutralization. The stronger binding of BLG despite its higher isoelectric point, inferred from lower pH(c), is shown to result from the negative "charge patch" on BLG, absent for BSA, as visualized via computer modeling (DelPhi). The higher affinity of BLG versus BSA was also confirmed by isothermal titration calorimetry (ITC). The relative values of pH(ϕ) for the two proteins show complex salt dependence so that the choice of ionic strength determines the order of coacervation, whereas the choice of pH controls the yield of the target protein. Coacervation at I = 100 mM, pH 7, of BLG from a 1:1 (w/w) mixture with BSA was shown by SEC to provide 90% purity of BLG with a 20-fold increase in concentration. Ultrafiltration was shown to remove effectively the polymer from the target protein. The relationship between protein charge anisotropy and binding affinity and between binding affinity and selective coacervation, inferred from the results for BLG/BSA, was tested using the isoforms of BLG. Substitution of glycine in BLG-B by aspartate in BLG-A lowers pH(c) by 0.2, as anticipated on the basis of DelPhi modeling. The stronger binding of BLG-A, confirmed by ITC, led to a difference in pH(ϕ) that was sufficient to provide enrichment by a factor of 2 for BLG-A in the coacervate formed from "native BLG".

Electrostatic selectivity in protein-nanoparticle interactions.

The binding of bovine serum albumin (BSA) and ß-lactoglobulin (BLG) to TTMA (a cationic gold nanoparticle coupled to 3,6,9,12-tetraoxatricosan-1-aminium, 23-mercapto-N,N,N-trimethyl) was studied by high-resolution turbidimetry (to observe a critical pH for binding), dynamic light scattering (to monitor particle growth), and isothermal titration calorimetry (to measure binding energetics), all as a function of pH and ionic strength. Distinctively higher affinities observed for BLG versus BSA, despite the lower pI of the latter, were explained in terms of their different charge anisotropies, namely, the negative charge patch of BLG. To confirm this effect, we studied two isoforms of BLG that differ in only two amino acids. Significantly stronger binding to BLGA could be attributed to the presence of the additional aspartates in the negative charge domain for the BLG dimer, best portrayed in DelPhi. This selectivity decreases at low ionic strength, at which both isoforms bind well below pI. Selectivity increases with ionic strength for BLG versus BSA, which binds above pI. This result points to the diminished role of long-range repulsions for binding above pI. Dynamic light scattering reveals a tendency for higher-order aggregation for TTMA-BSA at pH above the pI of BSA, due to its ability to bridge nanoparticles. In contrast, soluble BLG-TTMA complexes were stable over a range of pH because the charge anisotropy of this protein at makes it unable to bridge nanoparticles. Finally, isothermal titration calorimetry shows endoenthalpic binding for all proteins: the higher affinity of TTMA for BLGA versus BLGB comes from a difference in the dominant entropy term.

Boronic acid-functionalized spherical polymer brushes for efficient and selective enrichment of glycoproteins.

Glycoproteins are related to many biological activities and diseases, and thereby their efficient capture and enrichment for diagnostics and proteomics have emerged to exhibit great significance. However, the lack of materials with high binding capacity and selectivity is still a big obstacle for further application. Herein, we reported a facile and eco-friendly approach to fabricate spherical polymer brushes with multiple boronic acid groups. Specifically, the whole process can be divided into three steps, the polystyrene (PS) core was obtained by traditional emulsion polymerization, followed by immobilization of a home-made photoinitiator. Subsequently, boronic acid-functionalized polymer chains (PBA) were chemically grafted via photo-emulsion polymerization, leading to spherical polymer brushes (PS-PBA) with boronate affinity. The particle size, morphology, and composition of as-prepared spherical polymer brushes were systematically characterized. The characteristics of glycoproteins binding to the spherical polymer brushes under different conditions, including pH values and ionic strength, were also investigated. PS-PBA brushes possess fast binding speed (30 min) and high binding capacity for glycoprotein ovalbumin (OVA) (377.0 mg g-1) under physiological pH conditions at 25 °C, because the low steric hindrance of flexible polymeric PBA chains facilitates the interaction between boronic acid groups and glycoproteins. Moreover, the binding capacity of PS-PBA brushes for glycoprotein OVA was ∼6.7 times higher than that for non-glycoprotein bovine serum albumin (BSA), indicating the excellent selective adsorption. This study provided a facile and efficient approach for the fabrication of boronic acid-functionalized materials that will be useful in the enrichment and separation of glycoproteins for the diagnosis of diseases.

Recyclable spherical polyelectrolyte brushes containing magnetic nanoparticles in core.

Spherical polyelectrolyte brushes consisting of a magnetite/polystyrene nanocomposite core and a poly(acrylic acid) brush shell were prepared by photo-emulsion polymerization. They are narrowly dispersed, superparamagnetic and redispersible after aggregating by external magnetic field, as determined by transmission electron microscopy, dynamic light scattering, thermal gravimetric analysis and a vibrating sample magnetometer. Magnetic control is thus introduced into nano-sized spherical polyelectrolyte brushes to achieve recovery and controllable delivery in applications. This approach opens up the way for cost-effective applications of spherical polyelectrolyte brushes.

Conformation Variation and Tunable Protein Adsorption through Combination of Poly(acrylic acid) and Antifouling Poly(N-(2-hydroxyethyl) acrylamide) Diblock on a Particle Surface.

Adsorption and desorption of proteins on biomaterial surfaces play a critical role in numerous biomedical applications. Spherical diblock polymer brushes (polystyrene with photoiniferter (PSV) as the core) with different block sequence, poly(acrylic acid)-b-poly(N-(2-hydroxyethyl) acrylamide) (PSV@PAA-b-PHEAA) and poly(N-(2-hydroxyethyl) acrylamide)-b-poly(acrylic acid) (PSV@PHEAA-b-PAA) were prepared via surface-initiated photoiniferter-mediated polymerization (SI-PIMP) and confirmed by a series of characterizations including TEM, Fourier transform infrared (FTIR) and elemental analysis. Both diblock polymer brushes show typical pH-dependent properties measured by dynamic light scattering (DLS) and Zeta potential. It is interesting to find out that conformation of PSV@PAA-b-PHEAA uniquely change with pH values, which is due to cooperation of electrostatic repulsion and steric hindrance. High-resolution turbidimetric titration was applied to explore the behavior of bovine serum albumin (BSA) binding to diblock polymer brushes, and the protein adsorption could be tuned by the existence of PHEAA as well as apparent PAA density. These studies laid a theoretical foundation for design of diblock polymer brushes and a possible application in biomedical fields.

Hairy Fluorescent Nanospheres Based on Polyelectrolyte Brush for Highly Sensitive Determination of Cu(II).

Currently, it is an ongoing challenge to develop fluorescent nanosphere detectors that are uniform, non-toxic, stable and bearing a large number of functional groups on the surface for further applications in a variety of fields. Here, we have synthesized hairy nanospheres (HNs) with different particle sizes and a content range of carboxyl groups from 4 mmol/g to 9 mmol/g. Based on this, hairy fluorescent nanospheres (HFNs) were prepared by the traditional coupling method (TCM) or adsorption-induced coupling method (ACM). By comparison, it was found that high brightness HFNs are fabricated based on HNs with poly (acrylic acid) brushes on the surface via ACM. The fluorescence intensity of hairy fluorescent nanospheres could be controlled by tuning the content of 5-aminofluorescein (5-AF) or the carboxyl groups of HNs easily. The carboxyl content of the HFNs could be as high as 8 mmol/g for further applications. The obtained HFNs are used for the detection of heavy metal ions in environmental pollution. Among various other metal ions, the response to Cu (II) is more obvious. We demonstrated that HFNs can serve as a selective probe and for the separation and determination of Cu(II) ions with a linear range of 0-0.5 µM and a low detection limit of 64 nM.

Synthesis of poly(acrylic acid) coated magnetic nanospheres via a multiple polymerization route.

Magnetic nanospheres are versatile candidates for both fundamental and practical applications. Before they are applied in more complicated fields, their surface must be modified by several functionalities. However, the surface modification can be affected by the magnetic nanoparticles (MNP) embedded in the polymer matrix. Herein, the synthesis of poly(acrylic acid) coated magnetic nanospheres via a multiple polymerization route is described. During the synthesis process, seed emulsion polymerization was applied to redistribute the MNP in the polymer matrix, and the relationship between the structure of magnetic nanospheres and the thickness of the grafted poly(acrylic acid) layer was investigated. The development of size, morphology and magnetic properties of the nanospheres were characterized by transmission electron microscopy, dynamic light scattering, thermogravimetric analysis, X-ray diffraction and vibrating sample magnetometry. This work would pave the way to design and preparation of new structure of functional magnetic nanospheres with precise surface modification.

Stabilization of Pickering Emulsions by Hairy Nanoparticles Bearing Polyanions.

Pickering emulsions are increasingly applied in drug delivery, oil-water separation, composite materials preparation, and other fields. However, systematic studies on the stabilization of Pickering emulsions to satisfy the growing application demands in multiple fields with long-term conservation are rare. Compared to conventional solid nanoparticles, polyanion-modified hairy nanoparticles are more stable in practical environments and are investigated in this study. Poly (sodium p-styrenesulfonate) was grafted to a polystyrene (PS) core via a photoemulsion polymerization. A hairy nanoparticle bearing polyanions called poly (sodium p-styrenesulfonate) brush (PS@PSS) was synthesized. The size and uniformity of the Pickering emulsions stabilized by PS@PSS were investigated via a polarizing microscope. The stability of Pickering emulsions were optimized by adjusting critical factors like ultrasonic power and time, standing time, oil phases, salt concentration, and water:oil ratio. Results indicated that the Pickering emulsions could be stabilized by PS@PSS nanoparticles, which showed remarkable and adjustable partial wetting properties. It was found that the optimized conditions were ultrasonic power of 150 W, ultrasonic time of 3 min, salt concentration of 0.1 mM, oil phase of hexadecane, and water:oil ratio of 1:1. The formation and stability of Pickering emulsion are closely related to the hairy poly (sodium p-styrenesulfonate) brush layer on the nanoparticle surface.

Design and preparation of bi-functionalized short-chain modified zwitterionic nanoparticles.

An ideal nanomaterial for use in the bio-medical field should have a distinctive surface capable of effectively preventing nonspecific protein adsorption and identifying target bio-molecules. Recently, the short-chain zwitterion strategy has been suggested as a simple and novel approach to create outstanding anti-fouling surfaces. In this paper, the carboxyl end group of short-chain zwitterion-coated silica nanoparticles (SiO2-ZWS) was found to be difficult to functionalize via a conventional EDC/NHS strategy due to its rapid hydrolysis side-reactions. Hence, a series of bi-functionalized silica nanoparticles (SiO2-ZWS/COOH) were designed and prepared by controlling the molar ratio of 3-aminopropyltriethoxysilane (APTES) to short-chain zwitterionic organosiloxane (ZWS) in order to achieve above goal. The synthesized SiO2-ZWS/COOH had similar excellent anti-fouling properties compared with SiO2-ZWS, even in 50% fetal bovine serum characterized by DLS and turbidimetric titration. Subsequently, SiO2-ZWS/COOH5/1 was chosen as a representative and then demonstrated higher detection signal intensity and more superior signal-to-noise ratios compare with the pure SiO2-COOH when they were used as a bio-carrier for chemiluminescence enzyme immunoassay (CLEIA). These unique bi-functionalized silica nanoparticles have many potential applications in the diagnostic and therapeutic fields. STATEMENT OF SIGNIFICANCE: Reducing nonspecific protein adsorption and enhancing the immobilized efficiency of specific bio-probes are two of the most important issues for bio-carriers, particularly for a nanoparticle based bio-carrier. Herein, we designed and prepared a bi-functional nanoparticle with anti-fouling property and bio conjugation capacity for further bioassay by improving the short-chain zwitterionic modification strategy we have proposed previously. The heterogeneous surface of this nanoparticle showed effective anti-fouling properties both in model protein solutions and fetal bovine serum (FBS). The modified nanoparticles can also be successfully functionalized with a specific antibody for CLEIA assay with a prominent bio-detection performance even in 50% FBS. In this paper, we also investigated an unexpectedly fast hydrolysis behavior of NHS-activated carboxylic groups within the pure short-chain zwitterionic molecule that led to no protein binding in the short-chain zwitterion modified nanoparticle. Our findings pave a new way for the designing of high performance bio-carriers, demonstrating their strong potential as a robust platform for diagnosis and therapy.