High-throughput, low-cost quantification of 11 therapeutic antibodies using caprylic acid precipitation and LC-MS/MS.

BACKGROUND: Therapeutic drug monitoring of treatment with therapeutic antibodies is hampered by the application of a wide range of different methods in the quantification of serum levels. LC-MS based methods could significantly improve comparability of results from different laboratories, but such methods are often considered complicated and costly. We developed a method for LC-MS/MS based quantification of 11 therapeutic antibodies concomitantly measured in a single run, with emphasis on simplicity in sample preparation and low cost. RESULTS: After a single-step sample purification using caprylic acid precipitation to remove interfering proteins, the sample underwent proteolysis followed by LC-MS/MS analysis. Infliximab is used as internal standard for sample preparation while isotope-labeled signature peptides identified for each analyte are internal standards for the LC-MS/MS normalization. The method was validated according to recognized guidelines, and pipetting steps can be performed by automated liquid handling systems. The total precision of the method ranged between 2.7 and 7.3 % (5.1 % average) across the quantification range of 4-256 µg/ml for all 11 drugs, with an average accuracy of 96.3 %. Matrix effects were xamined in 55 individual patient samples instead of the recommended 6, and 147 individual patient samples were screened for interfering compounds. SIGNIFICANCE AND NOVELTY: Our method for simultaneous quantification of 11 t-mAb in human serum allows an unprecedented integration of robustness, speed and reduced complexity, which could pave the way for uniform use in research projects and clinical settings alike. In addition, the first LC-MS protocol for signature peptide-based quantification of durvalumab is described. This high throughput method can be readily adapted to a drug panel of choice.

Electron Microscopy for the Stability Assessment of Parenteral Nutrition Admixtures: Focus on Precipitation.

(1) Background: parenteral nutrition (PN) is indispensable for patients unable to receive oral or enteral feeding. However, the complexity of PN solutions presents challenges regarding stability and compatibility. Precipitation reactions may occur. The most frequent is the formation of calcium phosphate (Ca-P). The different factors influencing these reactions must be considered to ensure patient safety. (2) Methods: eight paediatric PN solutions were prepared, following standard protocols. Samples were stored at room temperature and in a refrigerator. Electron microscopy, coupled with energy dispersive X-ray spectroscopy (EDS), was employed. Precipitates were analysed for composition and morphology. (3) Results: precipitates were observed in all samples, even at day 0. Crystalline structures, predominantly composed of calcium or magnesium, sometimes associated with chlorine or phosphorus, were detected. Additionally, amorphous precipitates, contained heterogeneous compositions, including unexpected elements, were identified. (4) Conclusions: various precipitates, primarily calcium- or magnesium-based, can form in PN solutions, although it is not expected that they can form under the real conditions of use. Calcium oxalate precipitation has been characterised, but the use of organic calcium and phosphate salts appears to mitigate calcium phosphate precipitation. Electron microscopy provides interesting results on NP precipitation, but sample preparation may present technical limitations that affect the interpretation of the results.

Characterization of effective, simple, and low-cost precipitation methods for depleting abundant plasma proteins to enhance the depth and breadth of plasma proteomics.

Plasma is an abundant source of proteins and potential biomarkers to aid in the detection, diagnosis, and prognosis of human diseases. These proteins are often present at low levels in the blood and difficult to identify and measure due to the large dynamic range of proteins. The goal of this work was to characterize and compare various protein precipitation methods related to how they affect the depth and breadth of plasma proteomic studies. Abundant protein precipitation with perchloric acid (PerCA) can increase protein identifications and depth of plasma proteomic studies. Three acid- and four solvent-based precipitation methods were evaluated. All methods tested provided excellent plasma proteomic coverage (>600 identified protein groups) and detected protein in the low pg/mL range. Functional enrichment analysis revealed subtle differences within and larger changes between the precipitant groups. Methanol-based precipitation outperformed the other methods based on identifications and reproducibility. The methods' performance was verified using eight lung cancer patient samples, where >700 protein groups were measured and proteins with an estimated plasma concentration of ∼10 pg/mL were detected. Various protein precipitation agents are amenable to extending the depth and breadth of plasma proteomes. These data can guide investigators to implement inexpensive, high-throughput methods for their plasma proteomic workflows.

Microbially Induced Calcium Carbonate Precipitation by Sporosarcina pasteurii: a Case Study in Optimizing Biological CaCO3 Precipitation.

Current production of traditional concrete requires enormous energy investment that accounts for approximately 5 to 8% of the world's annual CO2 production. Biocement is a building material that is already in industrial use and has the potential to rival traditional concrete as a more convenient and more environmentally friendly alternative. Biocement relies on biological structures (enzymes, cells, and/or cellular superstructures) to mineralize and bind particles in aggregate materials (e.g., sand and soil particles). Sporosarcina pasteurii is a workhorse organism for biocementation, but most research to date has focused on S. pasteurii as a building material rather than a biological system. In this review, we synthesize available materials science, microbiology, biochemistry, and cell biology evidence regarding biological CaCO3 precipitation and the role of microbes in microbially induced calcium carbonate precipitation (MICP) with a focus on S. pasteurii. Based on the available information, we provide a model that describes the molecular and cellular processes involved in converting feedstock material (urea and Ca2+) into cement. The model provides a foundational framework that we use to highlight particular targets for researchers as they proceed into optimizing the biology of MICP for biocement production.

Design and preliminary techno-economic assessment of a pilot scale pharmaceutical wastewater treatment system for ammonia removal and recovery of fertilizer.

Recovery of nutrients from wastewater has a paramount importance for a sustainable and safe environment. In this study removal of ammonia and recovery of resources in the form of struvite from a complex pharmaceutical acidic wastewater having high concentration of ammoniacal nitrogen (NH4-N > 40 g/L) and other co-existing contaminants (magnesium, phosphorous, phenol etc.) was explored. Response Surface Methodology (RSM) was employed for design of experiments and process optimization. RSM results revealed that removal of ammoniacal nitrogen, i.e., struvite precipitation was found to be maximum in alkaline pH (10.5-11.0) at a N:Mg molar ratio (1:0.030 to 1:0.035) and N:P molar ratio (1:0.025 to 1:0.030). X-Ray diffraction, thermo-gravimetric analysis and Fourier transform-infrared spectroscopy confirmed the presence of struvite crystals in the obtained precipitate. Techno-economic assessment (TEA) based on mass energy balance principle and market equipment specifications revealed that a pilot-scale plant set up would have a break-even period of 1.06 years with a return on investment as 94.28%. This clearly elucidated the economic viability of the developed process for industrial applications for management of high ammonia laden pharmaceutical wastewater. While further specific technological improvements are needed for reduction of cost, this study will guide researchers and industries for careful selection of target markets to reduce the cost for successful implementation.

A comprehensive techno-economic analysis of income-generating sources on the conversion of real sheep slaughterhouse waste stream into valorized by-products.

The present analysis was conducted as the first research to assess the techno-economic viability of the value-added by-products (struvite, blood meal, bone meal, and raw sheepskin) from a medium-scale sheep slaughterhouse facility with a slaughtering capacity of 300 sheep per day. For this aim, a comparative technical and economic feasibility analysis was performed to assess the synergistic use of slaughterhouse-oriented rendering wastes and struvite recovery from real sheep abattoir effluent within the framework of detailed cost breakdown, break-even point, and payback period analyses. The experimental findings clearly showed that under the optimal conditions (chemical combination of MgCl2.6H2O + NaH2PO4.2H2O, a molar ratio of Mg2+:NH4+-N:PO43--P = 1.2:1:1, a reaction pH of 9.0, an initial ammonium concentration of 240 mg NH4+-N/L, and a reaction time of 15 min), struvite precipitation could effectively remove about 73%, 64%, 59%, and 82% of NH4+-N, TCOD, SCOD, and color, respectively, from the real sheep slaughterhouse waste stream. Based on various up-to-date techno-economic items considered within the break-even point analysis, the sheep slaughterhouse facility was estimated to achieve the targeted net income (€100/day) for any selling prices of €1041.30/ton, €640.05/ton, €263.72/ton, and €1.012/hide, respectively, for struvite, blood meal, bone meal, and raw sheepskin. Steel construction and chemicals were determined as the most costly components for CAPEX (capital expenditures) and OPEX (operating expenditures), respectively, and selling prices of bone meal and raw sheepskin were found to be the most critical income items on the profitability of the slaughterhouse facility. Co-monetary assessment of the struvite process and valorized compounds corroborated the economic viability of the proposed project with the payback periods of about 6.3 and 5.5 years, respectively, for the current market and the profit-oriented conditions without subsidy. The findings of this feasibility analysis, as the first of its own, could be used as guideline for simplifying the decision-making with regards to the feasibility of similar facilities and commercialization of profitable by-products.

Comparative assessment of the orthodontic wire's friction coated with zinc oxide nanoparticles by two methods of chemical precipitation and hydrothermal process.

INTRODUCTION: In orthodontic treatment with sliding technique, reduction of frictional forces could result in a more effective treatment. Recently, wire coating with nanoparticles were proposed to reduce frictional forces.

Strategies for the Removal of Polysaccharides from Biorefinery Lignins: Process Optimization and Techno Economic Evaluation.

The utilization of biorefinery lignins as a renewable resource for the production of bio-based chemicals and materials remain a challenge because of the high polysaccharide content of this variety of lignins. This study provides two simple methods; (i) the alkaline hydrolysis-acid precipitation method and (ii) the acid hydrolysis method for the removal of polysaccharides from polymeric biorefinery lignin samples. Both purification strategies are optimized for two different hardwood hydrolysis lignins, HL1 and HL2, containing 15.1% and 10.1% of polysaccharides, respectively. The treated lignins are characterized by polysaccharide content, molecular weight, hydroxyl content, and Attenuated Total Reflection-Fourier Transform Infrared Spectroscopy (ATR-FTIR). Preliminary techno-economic calculations are also carried out for both purification processes to assess the economic potential of these technologies. The results indicate that both protocols could be used for the purification of HL1 and HL2 hydrolysis lignins because of the minimal polysaccharide content obtained in the treated lignins. Nevertheless, from an industrial and economic perspective the acid hydrolysis technology using low acid concentrations and high temperatures is favored over the alkaline hydrolysis-acid precipitation strategy.

Simple, Inexpensive RNA Isolation and One-Step RT-qPCR Methods for SARS-CoV-2 Detection and General Use.

The most common method for RNA detection involves reverse transcription followed by quantitative polymerase chain reaction (RT-qPCR) analysis. Commercial one-step master mixes-which include both a reverse transcriptase and a thermostable polymerase and thus allow performing both the RT and qPCR steps consecutively in a sealed well-are key reagents for SARS-CoV-2 diagnostic testing; yet, these are typically expensive and have been affected by supply shortages in periods of high demand. As an alternative, we describe here how to express and purify Taq polymerase and M-MLV reverse transcriptase and assemble a homemade one-step RT-qPCR master mix. This mix can be easily assembled from scratch in any laboratory equipped for protein purification. We also describe two simple alternative methods to prepare clinical swab samples for SARS-CoV-2 RNA detection by RT-qPCR: heat-inactivation for direct addition, and concentration of RNA by isopropanol precipitation. Finally, we describe how to perform RT-qPCR using the homemade master mix, how to prepare in vitro-transcribed RNA standards, and how to use a fluorescence imager for endpoint detection of RT-PCR amplification in the absence of a qPCR machine In addition to being useful for diagnostics, these versatile protocols may be adapted for nucleic acid quantification in basic research. © 2021 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Preparation of a one-step RT-qPCR master mix using homemade enzymes Basic Protocol 2: Preparation of swab samples for direct RT-PCR Alternate Protocol 1: Concentration of RNA from swab samples by isopropanol precipitation Basic Protocol 3: One-step RT-qPCR of RNA samples using a real-time thermocycler Support Protocol: Preparation of RNA concentration standards by in vitro transcription Alternate Protocol 2: One-step RT-PCR using endpoint fluorescence detection.

Nitrogen recovery from pig slurry by struvite precipitation using a low-cost magnesium oxide.

Ammonia nitrogen management is a recurrent problem in intensive livestock areas. Struvite precipitation stands as a mature technology to recover ammonia nitrogen and prevent associated environmental problems. However, the feasibility of struvite technology to recover ammonia nitrogen from pig manure is limited by the reagents cost. This research aimed to optimise the formulation of a stabilizing agent (SA) synthesised using an industrial low-grade MgO by-product (LG-MgO) and phosphoric acid for efficient TAN recovery via struvite precipitation. Experimental results showed that the H3PO4/LG-MgO ratio controls the magnesium phosphate mineral phase of the SA (bobierrite and/or newberyite). Newberyite-rich SA showed the highest TAN removal efficiency from pig manure (66-73%) compared to the SA formed by a mixture of newberyite and bobierrite (51-59%) and by bobierrite (26%). Particle size reduction of LG-MgO did not improve the SA's TAN removal efficiency, although XRD patterns showed that the precipitates from the TAN removal experiments contained some unreacted newberyite. The economic analysis showed that the higher reactivity of the SA formulated using higher H3PO4/LG-MgO ratios compensated reagent costs. The SA synthesised with a H3PO4/LG-MgO ratio of 0.98 showed the most economical treatment cost, which was estimated at 7.5 € per kg of ammonia nitrogen from pig manure. Finally, the optimum SA was successfully synthesised in a 200-L pilot plant, with a TAN removal capacity only 10% lower than the one synthesised at lab-scale.

Stabilization of borate by hot isostatic pressing after co-precipitation with hydroxyapatite using MAP.

Boric acid is one of the most mobile inorganic contaminant species in nature due to its pKa of 9.23. Co-precipitation of borate with hydroxyapatite (HAp: Ca5(PO4)3OH) facilitates the simultaneous removal of borate with co-existing oxoanions in natural waters. The cost of phosphate is an impediment to industrialize the co-precipitation of borate with HAp for treatment of geothermal waters. In the present work, an inexpensive industrial by-product of magnesium ammonium phosphate (MAP) derived from sewage sludge, was examined as a phosphate source. MAP includes 89% pure magnesium ammonium phosphate, resulting in better performance than the pure chemical form of NH4H2PO4, because Mg2+ and Al3+ (trace elements in MAP product) play roles in enhancing the removal rate of borate and lowering the equilibrium borate concentration. These ions have a good affinity with phosphate to nucleate crystal seeds independently of powdery Ca sources. To reduce the bulky volume of solid residues, hot isostatic pressing (HIP) was applied. There is structural water in HAp; therefore, the greatest volume reduction was achieved with 78.3 ± 2.0% (n = 3). Additionally, a synergic effect to suppress the released borate, greater than the sequential combination of calcination and cold isostatic pressing was accomplished in the toxicity contents leaching procedure (TCLP) test. This is not due to larger crystal sizes alone, but it is derived from boron stabilization in HAp at an atomic level by the synergic effect of heating and pressing simultaneously.

Technical-economic comparison of chemical precipitation and ion exchange processes for the removal of phosphorus from wastewater.

Chemical precipitation with the addition of ferric chloride is commonly used to remove phosphorus from wastewater. However, since its application also involves several disadvantages, alternative solutions are required. The present paper shows the results of a full-scale experimental work aimed at evaluating the efficiency of the ion exchange process using a polymeric anion exchange resin impregnated with aluminum ions in the removal of phosphorus from wastewater. The study compared the results obtained through this process with chemical precipitation, considering both technical and economic issues. At the same dosage of 6 L/hour and influent concentration (about 6 mg/L), total removal efficiency of 95% and 78% (including also that occurring in the mechanical and biological processes) was achieved by means of the anion exchange process and chemical precipitation, respectively. However, in the latter case, this value was insufficient to ensure consistent compliance with the limit of 2 mg/L Ptot set on the effluent; to achieve this goal, the ferric chloride dosage had to be raised to 12 L/hour, thus increasing the related costs. Furthermore, the anion exchange process generated a lower sludge production. Therefore, the ion exchange process represents a valid alternative to chemical precipitation for P removal from wastewater.

Recent Advances in Co-processed APIs and Proposals for Enabling Commercialization of These Transformative Technologies.

Optimized physical properties (e.g., bulk, surface/interfacial, and mechanical properties) of active pharmaceutical ingredients (APIs) are key to the successful integration of drug substance and drug product manufacturing, robust drug product manufacturing operations, and ultimately to attaining consistent drug product critical quality attributes. However, an appreciable number of APIs have physical properties that cannot be managed via routes such as form selection, adjustments to the crystallization process parameters, or milling. Approaches to control physical properties in innovative ways offer the possibility of providing additional and unique opportunities to control API physical properties for both batch and continuous drug product manufacturing, ultimately resulting in simplified and more robust pharmaceutical manufacturing processes. Specifically, diverse opportunities to significantly enhance API physical properties are created if allowances are made for generating co-processed APIs by introducing nonactive components (e.g., excipients, additives, carriers) during drug substance manufacturing. The addition of a nonactive coformer during drug substance manufacturing is currently an accepted approach for cocrystals, and it would be beneficial if a similar allowance could be made for other nonactive components with the ability to modify the physical properties of the API. In many cases, co-processed APIs could enable continuous direct compression for small molecules, and longer term, this approach could be leveraged to simplify continuous end-to-end drug substance to drug product manufacturing processes for both small and large molecules. As with any novel technology, the regulatory expectations for co-processed APIs are not yet clearly defined, and this creates challenges for commercial implementation of these technologies by the pharmaceutical industry. The intent of this paper is to highlight the opportunities and growing interest in realizing the benefits of co-processed APIs, exemplified by a body of academic research and industrial examples. This work will highlight reasons why co-processed APIs would best be considered as drug substances from a regulatory perspective and emphasize the areas where regulatory strategies need to be established to allow for commercialization of innovative approaches in this area.

A Simple, Cost-Effective, and Robust Method for rRNA Depletion in RNA-Sequencing Studies.

The profiling of gene expression by RNA sequencing (RNA-seq) has enabled powerful studies of global transcriptional patterns in all organisms, including bacteria. Because the vast majority of RNA in bacteria is rRNA, it is standard practice to deplete the rRNA from a total RNA sample such that the reads in an RNA-seq experiment derive predominantly from mRNA. One of the most commonly used commercial kits for rRNA depletion, the Ribo-Zero kit from Illumina, was recently discontinued abruptly and for an extended period of time. Here, we report the development of a simple, cost-effective, and robust method for depleting rRNA that can be easily implemented by any lab or facility. We first developed an algorithm for designing biotinylated oligonucleotides that will hybridize tightly and specifically to the 23S, 16S, and 5S rRNAs from any species of interest. Precipitation of these oligonucleotides bound to rRNA by magnetic streptavidin-coated beads then depletes rRNA from a complex, total RNA sample such that ∼75 to 80% of reads in a typical RNA-seq experiment derive from mRNA. Importantly, we demonstrate a high correlation of RNA abundance or fold change measurements in RNA-seq experiments between our method and the Ribo-Zero kit. Complete details on the methodology are provided, including open-source software for designing oligonucleotides optimized for any bacterial species or community of interest.IMPORTANCE The ability to examine global patterns of gene expression in microbes through RNA sequencing has fundamentally transformed microbiology. However, RNA-seq depends critically on the removal of rRNA from total RNA samples. Otherwise, rRNA would comprise upward of 90% of the reads in a typical RNA-seq experiment, limiting the reads coming from mRNA or requiring high total read depth. A commonly used kit for rRNA subtraction from Illumina was recently unavailable for an extended period of time, disrupting routine rRNA depletion. Here, we report the development of a "do-it-yourself" kit for rapid, cost-effective, and robust depletion of rRNA from total RNA. We present an algorithm for designing biotinylated oligonucleotides that will hybridize to the rRNAs from a target set of species. We then demonstrate that the designed oligonucleotides enable sufficient rRNA depletion to produce RNA-seq data with 75 to 80% of reads coming from mRNA. The methodology presented should enable RNA-seq studies on any species or metagenomic sample of interest.

Optimization of bacterial sporulation using economic nutrient for self-healing concrete.

The use of heat- and alkali-resistant bacteria is essential for the biological repair of damaged concrete. Lysinibacillus boronitolerans YS11 was isolated from the rhizosphere of Miscanthus sacchariflorus. The increased pH in the urea-minus condition during the growth of the YS11 strain promoted calcium carbonate (CaCO3) formation. To identify the optimum medium that promoted the growth of the YS11 strain, a Plackett-Burman design was conducted for the screening process. Consequently, malt powder, rice bran, (NH4)2SO4, and corn syrup were chosen to enhance YS11 growth. The optimization of these four useful factors was carried out using a central composite design. To obtain higher survivability in mortar, the sporulation process is essential, and additional factors such as Mn2+, Fe2+, and Ca2+ were found to contribute to sporulation. A mixture of L. boronitolerans YS11 spore powder, cement, paste, sand, yeast extract, calcium lactate, and water showed a healing effect on a 0.3 mm mortar crack in 7 days. Furthermore, calcium carbonate precipitation was observed over the crack surface. Thus, we confirmed that mortar treated with YS11 spore powder was effective in healing micro-cracks in concrete.

Struvite crystallization by using raw seawater: Improving economics and environmental footprint while maintaining phosphorus recovery and product quality.

Seawater, as an alternative magnesium source, has the potential to improve the overall economics and environmental footprint of struvite production compared to the use of pure magnesium salts. However, the dilution effect and the presence of other ions in seawater can reduce the phosphorus recovery potential and the simultaneous precipitation of other compounds may reduce the quality of the produced struvite. This work presents a comparative study of seawater and MgCl2 by performing a series of thermodynamic equilibrium modeling and crystallization experiments. The results revealed that acceptable phosphorus recovery (80-90%) is achievable by using seawater as the magnesium source for struvite precipitation. Further, the simultaneous precipitation of calcium phosphates was successfully controlled and minimized by optimum selection of reaction pH and seawater volume (i.e. Mg:P and Mg:Ca molar ratios). The increase of temperature from 20 °C to 30 °C reduced the phosphorus recovery by 15-20% while it increased the particle size by 30-35%. The presence of suspended solids in reject water did not have significant effects on phosphorus recovery but it made the struvite separation difficult as the obtained struvite was mixed with suspended solids. The experimental results and economic evaluation showed that the use of seawater can reduce the chemical costs (30-50%) and the CO2-footprint (8-40%) of struvite production. It was concluded that seawater is a potential alternative to pure magnesium sources in struvite production, while studies in larger scale and continuous mode are needed for further verification before full-scale applications.

Simple and sensitive method for the analysis of 14 antituberculosis drugs using liquid chromatography/tandem mass spectrometry in human plasma.

Monitoring plasma concentration and adjusting doses of antituberculosis (TB) drugs are beneficial for improving responses to drug treatment and avoiding adverse drug reactions. A simple and sensitive liquid chromatography/tandem mass spectrometry method was developed to measure the plasma concentrations of 14 anti-TB drugs: ethambutol, isoniazid, pyrazinamide, levofloxacin, gatifloxacin, moxifloxacin, prothionamide, linezolid, rifampin, rifapentine, rifabutin, cycloserine, p-aminosalicylic acid, and clofazimine. METHODS: Human plasma was precipitated by acetonitrile and was subsequently separated by an AQ-C18 column with a gradient elution. Drug concentrations were determined using multiple reaction monitoring in positive ion electrospray ionization mode. According to pharmacokinetic data of patients, the peak concentration ranges and the timing of blood collection were determined. RESULTS: Intra- and interday precision was < 14.8%. Linearity, accuracy, extraction recovery, and matrix effect were acceptable for each drug. The stability of the method satisfied different storage conditions. CONCLUSIONS: The method allowed the sensitive and reproducible determination of 14 frequently used anti-TB drugs which has already been of benefit for some TB patients.

Enhanced electrochemical phosphate recovery from livestock wastewater by adjusting pH with plant ash.

In the field of environmental wastewater treatment, it is a very meaningful topic to recover phosphate from swine wastewater in the form of struvite precipitation. The solution pH is one of the important influencing factors in the process of struvite precipitation. In this paper, an attempt was made to recover the phosphate from swine wastewater by adding plant ash. Experimental results have revealed that aeration can be replaced by optimal plant ash adding mode to increase the phosphate recovery efficiency. With the dosages of plant ash and magnesium metal were respectively 11.66 and 3.33 g/L the phosphate recovery efficiency reached 97.69% in 60 min. The efficiency was still above 95% after repeatedly using magnesium pellet for 3 times. The economic evaluation further revealed that the recovery cost of the proposed method was 0.62 $/kg PO4-P.

Enhancing efficiency and economics of phosphorus recovery process by customizing the product based on sidestream characteristics - an alternative phosphorus recovery strategy.

The enhanced biological phosphorus removal process makes the phosphorus recovery feasible from the dewatering streams of biological sludge. The physicochemical properties of these sidestreams, as an input to a crystallizer, are different before and after anaerobic digestion. In this study, phosphorus recovery by calcium phosphate is proposed for pre-digestion sidestreams and by struvite precipitation for post-digestion sidestreams. The thermodynamic modeling followed by experimental tests was performed to evaluate the recovery efficiency and product properties of struvite and calcium phosphates. The variations in phosphorus recovery potential, reaction kinetics and particle size distribution emphasize the importance of the adjustment of initial supersaturation and pH of the reaction. The optimum pH, considering the economics and recovery efficiency, for both calcium phosphate and struvite precipitation was found to be pH = 8.5, whereas further increase of pH will not improve the overall efficiency of the process. In the case of calcium phosphate precipitation, it was shown that possible phase transformations should be considered and controlled as they affect both process efficiency and product properties. The economic evaluation indicated that the optimized operational condition should be determined for the phosphorus recovery process and that chemical costs for the production of calcium phosphates is lower than for struvite.