Human Low Density Lipoprotein (LDL) and Human Serum Albumin (HSA) Co‐Adsorption Onto the C18‐Silica Gradient Surface

Co‐adsorption kinetics of human low density lipoprotein (LDL) and serum albumin (HSA) on hydrophilic/hydrophobic gradient silica surface were studied using Total Internal Reflection Fluorescence (TIRF) and autoradiography. Two experimental systems were examined: (1) fluorescein‐labeled LDL (FITC‐LDL) adsorption from a FITC‐LDL + HSA solution mixture onto the octadecyldimethylsilyl (C18)‐silica gradient surface, and (2) the FITC‐LDL adsorption onto the HSA pre‐adsorbed on the C18‐silica gradient surface. Experiments with fluorescein‐labeled albumin (FITC‐HSA) and unlabeled LDL have been performed in parallel. The adsorption kinetics of FITC‐LDL onto the hydrophilic silica was found to be transport‐limited and not affected by co‐adsorption of HSA. A slower adsorption kinetics of lipoprotein onto the silica with pre‐adsorbed HSA layer resulted from a slow appearance of LDL binding sites exposed by the process of HSA desorption. In the region of increasing surface density of C18 groups, the FITC‐LDL adsorption rate fell below the transport‐limited adsorption rate, except in the very early adsorption times. Pre‐adsorption of HSA onto the C18‐silica gradient region resulted in a significant decrease of both the FITC‐LDL adsorption rate and adsorbed amount. The lowest FITC‐LDL adsorption was found in the region of C18 self‐assembled monolayer, where the pre‐adsorbed HSA layer almost completely eliminated lipoprotein binding.     

In-situ protein adsorption study on biofunctionalized surfaces using spectroscopic ellipsometry

Techniques currently employed to evaluate biomolecular interactions on surfaces require the use of radiolabeled, enzymatic, or fluorescent-tags to record and report the binding event. Ellipsometry has proven to be a powerful tool in understanding the biomolecular interactions on solid substrates and, typically does not require the labeling of the ligand or the receptor. In this present study, the adsorption kinetics of Human Serum Albumin (HSA) on functionalized silicon surfaces were evaluated using in-situ ellipsometry. In-situ ellipsometry was used to estimate the thickness of the adsorbed layers and the adsorption and desorption kinetics of HSA on functionalized surfaces. In this study, dense, self assembled monolayers were fabricated using aminopropyltriethoxysilane (APTES) and mixed silanes using APTES and methyltriethoxysilane at a ratio of 1:10, to serve as a template for protein immobilization on silicon surfaces. The silane derivatized surfaces were further modified using three different ligands/receptors that have been reported to bind HSA, namely: a linear peptide, a polyclonal antibody against human serum albumin, and small synthetic ligand (2, 4, 6-Tris(dimethylaminomethyl)phenol. The amount of HSA adsorbed was observed to increase with time, and with the initial concentration of the HSA solution. The adsorption kinetics of HSA on functionalized surfaces was approximated by a simple model for protein adsorption. A good model fit was obtained for the experimental data, thus enabling the interpretation of the adsorption kinetics of HSA on functionalized silicon surfaces. The effect of different HSA binding ligands on the rate constants affecting protein adsorption and desorption were studied.     

硅掺杂羟基磷灰石生物活性微粉对人血清白蛋白的吸附特性

对含硅羟基磷灰石(Si-HA)与人血清白蛋白(HSA)的吸附特性及动力学特征进行了研究,计算了表观活化能,用等温吸附曲线进行拟合。结果表明,该吸附属于Langmuir型。Si-HA和HA吸附HSA的表观活化能分别为21.28和35.57 kJ.mol-1,20℃下吸附反应的速率常数分别为1.48和1.34 min-1,饱和吸附量分别为25.34和21.34 mg.g-1,Si-HA具有比纯HA更好的吸附优势。通过XRD、FTIR及荧光光谱分析,探讨了HSA在Si-HA和HA表面的吸附作用机制,表明Si-HA与蛋白表面吸附反应是以化学吸附为主、包含物理吸附的混合吸附过程。从分子结构的角度揭示了Si-HA和HA吸附蛋白质性能的差异,为进一步探讨Si-HA和HA生物相容性提供了新途径。     

In situ ATR/FTIR studies of protein adsorption on polymeric materials: effectiveness of surface heparinization

The adsorption of two proteins from human plasma (human serum albumin (HSA) and human fibrinogen (HFG) onto six different polymeric surfaces (two of which are heparinized), has been studied byin situ ATR/FTIR spectroscopy. The different surface characteristics are reflected by different interfacial behaviours of the two proteins, but while both proteins unfold upon adsorption on all the different non-heparinized materials, they maintain the native conformation once adsorbed on the heparinized surfaces. These findings emphasize the effectiveness of surface heparinization.     

Preliminary study on human protein adsorption and leukocyte adhesion to starch-based biomaterials

In this study, the adsorption of human serum albumin (HSA), fibronectin (FN) and vitronectin (VN) onto the surface of novel biodegradable materials was evaluated by immunostaining. Specifically, polymeric blends of corn starch with cellulose acetate (SCA), ethylene vinyl alcohol copolymer (SEVA-C), and polycaprolactone (SPCL) were immersed in unitary and competitive systems; that is, binary and more complex protein solutions. For binary solutions, different HSA and FN protein distribution patterns were observed depending on the starch-based surface. Furthermore, the relative amount of proteins adsorbed onto starch-based surfaces was clearly affected by protein type: a preferential adsorption of VN and FN as compared to HSA was observed. On tests carried out with unitary, binary and more complex solutions, it was found that vitronectin adsorption ability was enhanced in competitive systems, which was associated with a lower amount of adsorbed albumin. In order to assess the effect of these human proteins on cell behavior, a mixed population of human lymphocytes and monocytes/macrophages was cultured over pre-coated SEVA-C surfaces. Through anti-CD3 and CD-14 monoclonal antibody labeling and cell counting, leukocyte adhesion onto pre-coated SEVA-C surfaces was analyzed. Based on the results, it was possible to detect albumin long-term effects and fibronectin short-term effects on cell adhesion proving that previously adsorbed proteins modulate leukocyte behavior.     

In Situ ATR‐FTIR Spectroscopy on the Deposition and Protein Interaction of Polycation/Alginate Multilayers

We report in situ ATR‐FTIR spectroscopy studies of the deposition of biocompatible polyelectrolyte multilayers (PEMs) consisting of the polycation chitosan (CHT) or poly(ethyleneimine) (PEI) and the polyanion sodium alginate (ALG) and their interactions with model‐protein human serum albumin (HSA). HSA‐adsorption data for PEI/ALG and CHT/ALG PEMs show the importance of the outermost polyelectrolyte (PEL) layer for protein interactions: CHT‐ or PEI‐terminated PEMs are highly attractive to HSA, while ALG‐terminated PEMs are repulsive, relevant for the generation of protein‐active or protein‐inert biomedical surfaces. The bound HSA is found to be located at the surface rather than the inner region of the PEM.     

Resistance to nonspecific protein adsorption by poly(vinyl alcohol) thin films adsorbed to a poly(styrene) support matrix studied using surface plasmon resonance.

Thin films of poly(vinyl alcohol) (PVA) polymer were prepared on a flat, nonporous, poly(styrene) support matrix by adsorption from aqueous solution and were characterized in order to investigate the nonspecific adsorption of proteins to a chromatographically relevant surface. The integrity and surface coverage of the PVA thin films were established by surface analysis and atomic force microscopy imaging. The adsorption of the PVA polymers to the poly(styrene) substrate and the nonspecific adsorption of proteins to the PVA-coated surface were monitored using surface plasmon resonance. PVA was strongly bound to the poly(styrene) surface, but the surface density of the adsorbed PVA polymers was affected substantially by the concentration, molecular weight, and degree of hydrolysis of PVA polymers used. There was evidence of increasing degrees of unfolding of the PVA polymer onto the poly(styrene) surface as the concentration of the the PVA coating solution increased. Complete PVA coverage of the poly(styrene) surface was observed at PVA concentrations of 0.1 mg/mL or greater but with significant influence of both molecular weight and degree of hydrolysis of the PVA polymers. Resistance of the PVA-coated poly(styrene) surface to the nonspecific adsorption of human serum albumin (HSA) correlated with the degree of surface coverage of the PVA. The use of anti-HSA as a probe for adsorbed HSA suggested that HSA was displacing PVA from the poly(styrene) surface at the lower PVA surface coverage. A complete barrier to nonspecific protein adsorption was observed with a PVA coating solution concentration of greater than 0.1 mg/ mL with a degree of hydrolysis of <88%.     

Use of piezoelectric-excited millimeter-sized cantilever sensors to measure albumin interaction with self-assembled monolayers of alkanethiols having different functional headgroups

In this paper, we describe a new modality of measuring human serum albumin (HSA) adsorption continuously on CH3-, COOH-, and OH-terminated self-assembled monolayers (SAMs) of C11-alkanethiols and the direct quantification of the adsorbed amount. A gold-coated piezoelectric-excited millimeter-sized cantilever (PEMC) sensor of 6-mm2 sensing area was fabricated, where resonant frequency decreases upon mass increase. The resonant frequency in air of the detection peak was 45.5 +/- 0.01 kHz. SAMs of C11-thiols (in absolute ethanol) with different end groups was prepared on the PEMC sensor and then exposed to buffer solution containing HSA at 10 microg/mL. The resonant frequency decreased exponentially and reached a steady-state value within 30 min. The decrease in resonant frequency indicates that the mass of the sensor increased due to HSA adsorption onto the SAM layer. The frequency change obtained for the HSA adsorption on CH3-, COOH-, and OH-terminated SAM were 520.8 +/- 8.6 (n = 3), 290.4 +/- 6.1 (n = 2), and 210.6 +/- 8.1 Hz (n = 3), respectively. These results confirm prior conclusions that albumin adsorption decreased in the order, CH(3) > COOH > OH. Observed binding rate constants were 0.163 +/- 0.003, 0.248 +/- 0.006, and 0.381 +/- 0.001 min(-1), for methyl, carboxylic, and hydroxyl end groups, respectively. The significance of the results reported here is that both the formation of self-assembled monolayers and adsorption of serum protein onto the formed layer can be measured continuously, and quantification of the adsorbed amount can be determined directly.     

棒状纳米微晶纤维素诱导制备花状TiO2纳米晶体

以棒状纳米微晶纤维素(Nanocrystalline cellulose,NCC)为形貌诱导剂,TiCl4为原料,采用水解法在70℃的温度下反应4～6 h,制备了TiO2花状纳米晶体.采用TEM、HRTEM、XRD和FTIR对不同条件下制得的TiO2晶体的微观形貌、晶粒尺寸和晶相组成进行了表征,探讨了TiCl4的用量及反应时间对晶体形貌和晶型的影响,并对其形成机理进行分析.纳米微晶纤维素表面富含大量羟基,可与TiO2之间形成氢键连接,促使TiO2在其表面的异质成核和生长,同时纳米微晶纤维素在TiO2表面的吸附作用,改变了各晶面的表面能和生长速度,使TiO2沿着[110]方向优先生长形成一维针状物,针状物再进一步聚集形成花状聚集体.以甲基橙为目标污染物,测试了所得TiO2纳米花状晶体的光催化性能.结果表明,随着TiCl4用量的增多及反应时间的延长,所制得的TiO2纳米晶体中金红石含量增多,形貌逐渐复杂化,光催化性能下降.     

Surface modifying of microporous PTFE capillary for bilirubin removing from human plasma and its blood compatibility

In this study, human serum albumin (HSA) was covalently immobilized onto the inner surface of microporous poly(tetrafluoroethylene) (MPTFE) capillaries for direct bilirubin removal from human plasma. To obtain active binding sites for HSA, the MPTFE capillaries were chemically functionalized by using a coating of poly(vinyl alcohol) (PVA)–glycidyl methacrylate (GMA) copolymers. Characterization of grafted MPTFE capillaries was verified by XPS, Fourier transform infrared spectroscopy (FT-IR), scanning electronic microscopy (SEM). Non-specific adsorption on the PVA–GMA coated capillary remains low (< 0.38 mg bilirubin/g), and higher affinity adsorption capacity, of up to 73.6 mg bilirubin/g polymer was obtained after HSA is immobilized. Blood compatibility of the grafted MPTFE capillary was evaluated by SEM and platelet rich plasma (PRP) contacting experiments. The experimental data on blood compatibility indicated that PVA-coated and PVA–GMA–HSA coated PTFE capillary showed a sharp suppress on platelets adhesion. The proposed method has the potential of serving in bilirubin removal in clinical application.     

Characterization of Alizarin Red S binding sites and structural changes on human serum albumin: a biophysical study

Alizarin Red S (ARS), is a water-soluble, widely used anthraquinone dye synthesized by sulfonation of alizarin. In this report, the binding of ARS to human serum albumin (HSA) was characterized by employing fluorescence, UV/vis absorption, circular dichroism (CD), and molecular modeling methods. The data of fluorescence spectra displayed that the binding of ARS to HSA is the formation of HSA-ARS complex at 1:1 stoichiometric proportion. Hydrophobic probe 8-anilino-1-naphthalenesulfonic acid (ANS) was employed and elucidated that the dye was located in subdomain IIIA. This phenomenon corroborates the result of site-specific probe displacement experiments, which demonstrate the dye is at indole-benzodiazepine site (Sudlow's site II); and it is also consistent with guanidine hydrochloride (GuHCl) induced HSA unfolding studies and molecular modeling simulations. The features of the dye, which led to structural perturbations of HSA, have also been studied in detail by methods of UV/vis, CD and three-dimensional fluorescence spectroscopy.     

Albumin adsorption on Cibacron Blue F3G-A immobilized onto oligo(ethylene glycol)-terminated self-assembled monolayers

Self-assembled monolayers can be tailored with specific ligands to a certain protein and at the same time prevent the non-specific adsorption of other proteins. Cibacron Blue F3G-A (CB-thiol) was successfully immobilized onto tetra(ethylene glycol)-terminated alkanethiol (CB-thiol). The affinity of human serum albumin (HSA) to immobilized Cibacron Blue F3G-A was studied using mixed thiolate self-assembled monolayers on gold with different n-alkyl chain lengths and functional terminal groups (CH₃-; OH- and tetra(ethylene glycol)). Surfaces were characterized using X-ray photoelectron spectroscopy and water contact angle measurements. Albumin adsorption and exchangeability of the adsorbed albumin molecules with other albumin molecules in solution were evaluated using ¹²⁵I-radiolabeled HSA. Competitive adsorption between albumin and fibrinogen to the different self-assembled monolayers (SAMs) was also investigated. Results showed that the incorporation of CB-thiol on the monolayers does not increase the HSA adsorption and reversibility on the SAMs. However, although specific adsorption of HSA to the immobilized Cibacron Blue F3G-A was not demonstrated, the presence of CB-thiol decreases the affinity of fibrinogen to the OH-terminated SAMs.     

The competitive adsorption of human proteins onto natural-based biomaterials

This study aims to further the understanding of nanoscale structures relevant for cellular recognition on contact and interaction with natural-based materials. The correlation between surface characteristics and protein adsorption from unitary and complex protein systems was investigated with respect to altering the bulk chemistry of the substrate material. Polymeric blends of starch and cellulose acetate, polycaprolactone (SPCL) and ethylene vinyl alcohol (SEVA-C) were used. Different proteins, bovine serum albumin, human serum albumin (HSA) and human fibronectin (HFN), were selected for this study. The construction of adsorption isotherms is an important starting point towards characterizing the interactions between surfaces and proteins. In this study, albumin adsorption isotherms fit the Freundlich model and were correlated with the chemistry and morphology of surfaces. In addition, protein distribution, quantification and competition were measured using fluorimetry and visualized by confocal microscopy. The analysis of unitary systems demonstrated that the adsorption of HSA was generally lower than that of HFN. In the latter case, SPCL and SEVA-C blends reached adsorption values of 97 and 89 per cent, respectively. In studying the co-adsorption of proteins, an increase in both HSA and HFN on SEVA-C surfaces was observed. SPCL showed no substantial increase in the adsorption of the proteins in competitive conditions. The similarity of these materials with other polysaccharide-based materials increases the relevance of the presented results. This study provides valuable information for the development of strategies towards the control of protein orientation and functionality as the availability of cell signalling epitopes for a broader family of materials that continue to be a significant component of this field of research.     

Effect of negative functionalisation of gold nanorods on conformation and activity of human serum albumin

The theranostic applications of gold nanorods (AuNRs) are limited due to the presence of cytotoxic cetrimonium bromide (CTAB) stabiliser, leading to the instigation of alternate stabilisers like negatively charged polystyrene sulphonate (PSS). Despite previous reports suggesting the impact of PSS‐AuNRs on cells, their effect on the most abundant protein in plasma, i.e. human serum albumin (HSA), has not been studied before. Hence, the impact of PSS‐AuNRs on HSA was thoroughly examined using varied spectroscopic techniques. The absorbance and fluorescence spectroscopic findings suggested the extent of ground‐state complexation and tryptophan domain disruptions of HSA for different AuNR concentrations, which were also suggested based on size measurements and activation energy calculations for complex formation. Modifications in the hydrophobic environment of HSA were evaluated using synchronous fluorescence, whereas the secondary structural damages were explained using circular dichroism (CD) and FTIR analyses. Additional studies to analyse protein denaturation, fibrillation, esterase activity, and free thiol were carried out to understand structural and functional changes. The study suggested that PSS‐AuNRs showed concentration‐dependent alterations in HSA structure, but the extent of protein toxicity was considerably lesser for PSS‐AuNRs of similar dimensions compared to the data available for CTAB‐AuNRs; thus, highlighting that PSS‐AuNRs could be safer for biomedical applications.Inspec keywords: fluorescence, gold, nanomedicine, molecular biophysics, hydrophobicity, cellular biophysics, circular dichroism, nanorods, biomedical materials, enzymes, Fourier transform infrared spectraOther keywords: HSA structure, PSS‐AuNRs, CTAB‐AuNRs, gold nanorods, human serum albumin, cytotoxic cetrimonium bromide stabiliser, negatively charged polystyrene sulphonate, absorbance, fluorescence spectroscopic findings, size measurements, activation energy calculations, AuNR concentrations, ground‐state complexation, tryptophan domain disruptions, hydrophobic environment, secondary structural damages, circular dichroism, FTIR analyses, protein denaturation, fibrillation, esterase activity, free thiol, concentration‐dependent alterations, protein toxicity     

Probing molecular interactions in bone biomaterials: Through molecular dynamics and Fourier transform infrared spectroscopy

Polymer-hydroxyapatite (HAP) composites are widely investigated for their potential use as bone replacement materials. The molecular interactions at mineral polymer interface are known to have significant role of mechanical response of the composite system. Modeling interactions between such dissimilar molecules using molecular dynamics (MD) is an area of current interest. Molecular dynamics studies require potential function or force field parameters. Some force fields are described in literature that represents the structure of hydroxyapatite reasonably well. Yet, the applicability of these force fields for studying the interaction between dissimilar materials (such as mineral and polymer) is limited, as there is no accurate representation of polymer in these force fields. We have obtained the parameters of consistent valence force field (CVFF) for monoclinic hydroxyapatite. Validation of parameters was done by comparing the computationally obtained unit cell parameters, vibrational spectra and atomic distances with XRD and FTIR experiments. Using the obtained parameters of HAP, and available parameters of polymer (polyacrylic acid), interaction study was performed with MD simulations. The MD simulations showed that several hydrogen bonds may form between HAP and polyacrylic acid depending upon the exposed surface of HAP. Also there are some favourable planes of HAP where polyacrylic acid is most likely to attach. We have also simulated the mineralization of HAP using a “synthetic biomineralization”. These modeling studies are supported by photoacoustic spectroscopy experiments on both porous and non porous composite samples for potential joint replacement and bone tissue engineering applications.     

Fabrication of a nanocarrier system through self-assembly of plasma protein and its tumor targeting

Human serum albumin (HSA) nanoparticles hold great promise as a nanocarrier system for targeted drug delivery. The objective of this study was to explore the possibility of preparing size controllable albumin nanoparticles using the disulfide bond breaking reagent β-mercaptoethanol (β-ME). The results showed that the protein concentration and temperature had positive effects on the sizes of the albumin nanoparticles, while pH had a negative effect on the rate of nanoparticle formation. The addition of β-ME induced changes in HSA secondary structure and exposed the hydrophobic core of HSA, leading to the formation of nanoparticles. Human serum albumin nanoparticles could be internalized by MCF-7 cells and mainly accumulated in cytoplasm. After injection in tumor bearing mice, the HSA nanoparticles accumulated in tumor tissues, demonstrating the targeting ability of the nanoparticles. Therefore, human serum albumin can be fabricated into nanoparticles by breaking the disulfide bonds and these nanoparticles exhibit high tumor targeting ability. Human serum albumin nanoparticles could be ideal for the targeted delivery of pharmacologically active substances.     

An X‐ray Spectromicroscopy Study of Albumin Adsorption to Crosslinked Polyethylene Oxide Films

Synchrotron‐based X‐ray photoemission electron microscopy (X‐PEEM) is used to characterize the near surface composition of polyethylene oxide (PEO) combined with 1.5, 5, and 10 wt.‐% pentaerythritol triacrylate (PETA) crosslinker. It is found that as the concentration of PETA increases, it becomes the dominant component in the top 10 nm of the film surface. The same surfaces are also exposed to human serum albumin (HSA) and the distributions of the protein relative to PEO and PETA measured with X‐PEEM. A positive correlation is found between levels of PETA and HSA at the surface. Above PETA concentrations of 5 wt.‐%, HSA adsorption is significant, which suggests high levels of PETA (often used to immobilize PEO by crosslinking) can significantly reduce the non‐fouling properties of PEO.     

Molecular modelling of protein adsorption on the surface of titanium dioxide polymorphs

This paper reports a molecular modelling study of the adsorption of protein subdomains with unlike secondary structures on different surfaces of ceramic titanium dioxide (TiO(2)), forming a passivating film on titanium biomaterials that provides the interface between the bulk metal and the physiological environment, affecting its biocompatibility and performance. Using molecular dynamics methods, we study the effect of the nanoscale structure of the common TiO(2) polymorphs (rutile, anatase and brookite) on the adsorption of an albumin subdomain and on two connected fibronectin modules, respectively containing α-helices and β-sheets. We find that the larger protein subdomain shows a stronger adsorption, as expected because of its size, but also that the three surfaces behave differently. In particular, brookite shows the weakest adsorption, whereas anatase leads to the strongest intrinsic adsorption, in particular for the fibronectin modules. Moreover, the simulations indicate a significant conformational change of the adsorbed protein subdomains with extensive surface nanopatterning. These results show that classical molecular dynamics methods can provide useful information about the influence of nanostructure and topology on protein physisorption at a fixed surface chemistry.     

Analysis of free drug fractions by ultrafast immunoaffinity chromatography

A chromatographic method was developed for measuring the nonbound (or free) fraction of drugs by using millisecond-scale extractions on small immunoaffinity columns. The design of this system was developed by considering the dissociation rates of (R)- and (S)-warfarin from the binding protein human serum albumin (HSA) and by performing computer simulations of the immunoaffinity extraction of these drugs. The final system was tested by using it to measure the free fractions of (R)- or (S)-warfarin in samples with known concentrations of these agents and HSA. The free warfarin fraction was extracted in 180 ms by a 2.1-mm-i.d. sandwich microcolumn that contained a 1.1-mm layer of an anti-warfarin antibody support. The nonretained peaks from this immunoaffinity column were passed through a series internal surface reversed-phase columns and a fluorescence detector for the analysis of any protein-bound warfarin that remained in the sample. The experimental results were found to have good agreement with those predicted from the known equilibrium constants for the binding of (R)- and (S)-warfarin with HSA. This approach can be modified for other analytes by changing the types of antibodies that are used in the immunoaffinity column and by using an appropriate detector for the nonretained drug fraction.     

Removal of phosphorus from aqueous solution by Posidonia oceanica fibers using continuous stirring tank reactor

The present study aims to develop a new potentially low-cost, sustainable treatment approach to soluble inorganic phosphorus removal from synthetic solutions and secondary wastewater effluents in which a plant waste (Posidonia oceanica fiber: POF) is used for further agronomic benefit. Dynamic flow tests using a continuous stirred tank reactor (CSTR) were carried out to study the effect of initial concentration of phosphorus, amount of adsorbent, feeding flow rate and anions competition. The experimental results showed that the removal efficiency of phosphorus from synthetic solutions is about 80% for 10 g L(-1) of POF. In addition, the variation of the initial concentration of phosphorus from 8 to 50 mg L(-1) increased the adsorption capacity from 0.99 to 3.03 mg g(-1). The use of secondary treated wastewater showed the presence of competition phenomenon between phosphorus and sulphate which could be overcoming with increasing the sorptive surface area and providing more adsorption sites when increasing the adsorbent dosage of POF. Compared with columns studies, this novel CSTR system showed more advantages for the removal of soluble phosphorus as a tertiary treatment of urban secondary effluents with more adsorption efficiency and capacity, in addition to the prospect use of saturated POF with nutriment as fertilizer and compost.