Marco Rito-Palomares

• Doctor of Engineering
• Head of Faculty at Tecnológico de Monterrey

Introduction Disulfiram (DSF) reduces insulin resistance and weight gain in obese mice. However, the effect on adipose tissue is unexplored due to their high instability under physiological conditions, limiting clinical applications. Thus, it is meaningful to develop a DSF carrier for sustained release to adipose tissue. We optimized the synthesis...

Obesity is a global disease characterized by excessive lipid accumulation in the adipose tissue. There is an urgent need to explore alternative compounds to treat obesity. Low- molecular-weight compounds from plants, like 3,3′-diindolylmethane (DIM), are emerging as potential alternatives for obesity treatment. In this work, DIM is encapsulated int...

Animal venoms are natural products that have served as a source of novel molecules that have inspired novel drugs for several diseases, including for metabolic diseases such as type-2 diabetes and obesity. From venoms, toxins such as exendin-4 (Heloderma suspectum) and crotamine (Crotalus durissus terrificus) have demonstrated their potential as tr...

Crotamine (Ctm) is a peptide isolated from Crotalus durissus terrificus venom. This molecule has been demonstrated to diminish body weight gain and enhance browning in adipose tissue, glucose tolerance, and insulin sensitivity; hence, it has been postulated as an anti-obesogenic peptide. However, the mechanism to elicit the anti-obesogenic effects...

BACKGROUND Rattlesnake venoms are a complex source of different toxin families with biotechnological and biomedical applications. However, the complexity of the sample and the low abundance of some toxins can restrict their study and applications. Toxin isolation usually requires multiple chromatographic steps, implying elevated operative costs and...

BACKGROUND Cell‐based high‐throughput platforms using aqueous two‐phase systems (ATPS) have led to the construction of 3D cultures using ATPS in which stem cells are confined into polymer–polymer droplets, giving rise to viable spheroids for their further proliferation and differentiation. This work aims to characterize polymer–polymer ATPS droplet...

Aqueous two-phase systems (ATPS) involve the combination of two immiscible polymers that at certain concentrations form a biphasic separation system. Due to the physicochemical properties and the low interfacial tension of the ATPS-forming polymers, the generation of ATPS droplets began to be explored. This work aims to characterize the generation...

Chemiluminescence (CL) reactions are widely used for the detection and quantification of many types of analytes. Laccase has previously been proposed in CL reactions; however, its light emission behavior has not been characterized. This study was conducted to characterize the laccase-luminol system, determine its kinetic parameters, and analyze the...

Purpose of Review Diabetes mellitus is a complex, chronic illness characterized by elevated blood glucose levels that occurs when there is cellular resistance to insulin action, pancreatic β-cells do not produce sufficient insulin, or both. Diabetes prevalence has greatly increased in recent decades; consequently, it is considered one of the fastes...

Abstract: A point-of-care (POC) can be defined as an in vitro diagnostic test that can provide results within minutes. It has gained enormous attention as a promising tool for biomarkers detection and diagnosis, as well as for screening of chronic noncommunicable diseases such as diabetes mellitus. Diabetes mellitus type 2 is one of the metabolic d...

Cattle tick (Rhipicephalus microplus) represents a severe problem causing substantial economic losses, estimated in billions of dollars annually. Currently, chemical acaricides represent the most widely used control method. However, several problems such as resistance have been described. Phage-based vaccines represent a fast and low-cost tool for...

Early detection is a key factor in patient fate. Currently, multiple biomolecules have been recognized as biomarkers. Nevertheless, their identification is only the starting line on the way to their implementation in disease diagnosis. Although blood is the biofluid par excellence for the quantification of biomarkers, its extraction is uncomfortabl...

Aqueous two‐phase systems (ATPS) have been demonstrated to be a simple and robust worldwide applied bioprocessing technology. From high value microscopic to macroscopic biological molecules obtained from a wide broad of matrices, ATPS have shown a high efficiency capacity for their concentration, enrichment, recovery, and purification. Particularly...

Bioseparation is a crucial process for the obtention of biological products that are essential in daily life, such as medicine and food. At first, the researchers centered their efforts on the optimization of the upstream processes, conducting to high product titer. Afterwards, they turned their attention to improving the downstream processes. Sinc...

The cover image is based on the Review Stem cell culture media enriched with plant-derived compounds: Cell proliferation enhancement by Alonso Ornelas-González et al., https://doi.org/10.1002/jctb.6794.

Proteins, which have inherent biorecognition properties, have long been used as therapeutic agents for the treatment of a wide variety of clinical indications. Protein modification through covalent attachment to different moieties improves the therapeutic’s pharmacokinetic properties, affinity, stability, confers protection against proteolytic degr...

The development of cell therapies to replace lost or damaged cells has attracted interest in the regenerative medicine field due to the multilineage differentiation ability of stem cells. However, there are some drawbacks regarding optimization in the differentiation protocols to obtain high efficiency and further translation into clinic settings,...

Stem cells are characterized by their self‐renewal and differentiation potential, emerging as a promising strategy for developing cell therapies to treat degenerative diseases. Stem cell culture under in vitro conditions involves the addition of growth factors to the media to stimulate their proliferation. However, most of these growth factors are...

Recently, stem cell-based therapies have been proposed as an alternative for the treatment of many diseases. Stem cells (SCs) are well known for their capacity to preserve themselves, proliferate, and differentiate into multiple lineages. These characteristics allow stem cells to be a viable option for the treatment of diverse diseases. Traditional...

Aqueous two-phase systems (ATPS) offer great opportunities regarding the purification of stem cells for future medical applications. However, most publications exploring the use of ATPS for the biopurification of stem cells are focused only on the use of a single system or in the comparison of different kinds of systems. These studies do not consid...

Xylanases are enzymes that degrade the linear polysaccharide xylan into xylose, thus breaking down hemicellulose, one of the major components of plant cell walls. Xylanases are applied in a wide range of industrial processes, such as in the pulp and paper industry, which demands a higher purity degree compared to other industries. Processes for the...

The increasing interest of the biopharmaceutical industry to exploit plants as a commercially viable production system is demanding the development of new strategies to maximize product recovery. Aqueous two-phase systems (ATPSs) are a primary recovery technique that has shown great potential for the efficient extraction and purification of biologi...

BACKGROUND The central stage of biotechnological production is the transformation of renewable raw materials into bioproducts in a water‐based bioreactor. The fermentation broth usually has a low concentration (less than 10%) of the bioproduct, which requires the application of subsequent separation and purification processes. For many bioproducts,...

Stem cell therapy has emerged as a promising alternative for replacing lost cells involved in neurodegenerative diseases. High efficiency of differentiation and full cell viability are actual challenges to achieve the translation of cell therapies to the clinic. To address this, the construction of aqueous two‐phase systems in three‐dimensional (AT...

Vaccines are considered one of the most important bioproducts in medicine. Since the development of the smallpox vaccine in 1796, several types of vaccines for many diseases have been created. However, some vaccines have shown limitations as high cost and low immune responses. In that regard, bacteriophages have been proposed as an attractive alter...

BACKGROUND Bacteriophages are bionanoparticles with several applications in different biotechnology‐based products. Among them, vaccines have the potential to treat antibiotic‐resistant bacteria and parasitic infections. Traditional methods for their recovery and purification rely on precipitation with polyethylene glycol (PEG) and NaCl. However, t...

Green fluorescent protein (GFP) is a useful biomolecule in biotechnology; however, its price makes its widespread application prohibitive. To overcome this issue, recently, the use of aqueous two-phase systems (ATPS) for GFP purification was proposed as an alternative platform to reduce processing costs. Aligned with this goal, this study performed...

Sedum dendroideum, a traditional medicinal plant, has a wide variety of pharmacological activities, attributed to phenolic compounds. Despite that there are several methods available for the extraction of phenolic compounds, their use has several disadvantages, evidencing the need for the development of alternative methods. In this study, ethanol-s...

The cover image is based on the Original Article Ribonuclease A modification with poly[N‐(2‐hydroxypropyl)methacrylamide] copolymers: new route of synthesis and purification by Marco Rito‐Palomares, Calef Sanchz‐Trasviña, and Karla Mayolo‐Deloisa., https://doi.org/10.1002/jctb.6367. The cover image is based on the Original Article Ribonuclease A mo...

Although microRNAs (miRNAs) have emerged as increasingly important target analytes, their biorecognition remains challenging due to their small size, high sequence homology, and low abundance in clinical samples. Nanospheres and microspheres have also gained increasing attention in biosensor applications due to their high specific surface area and...

CaptoTM Core 700 is a core-shell chromatographic support with an adsorbing core contained within an inert shell layer designed to purify larger biomolecules and bioparticles in a flow-through mode. The present study aims to characterize the structure and functional properties of this resin using bovine serum albumin (BSA, Mr∼65 kDa) and thyroglobul...

Laccase is a multi-copper oxidase that catalyzes the oxidation of one electron of a wide range of phenolic compounds. The enzyme is considered eco-friendly because it requires molecular oxygen as co-substrate for the catalysis and it yields water as the sole by-product. Laccase is commonly produced by fungi but also by some bacteria, insects and pl...

BACKGROUND N‐(2‐Hydroxypropyl)methacrylamide (HPMA), a synthetic polymer, has biocompatible features that allow it to be used as a drug carrier and used in drug delivery systems; however, its potential as a protein drug carrier has not been completely exploited. This study provides a new conjugation pathway to obtain HPMA–protein conjugates and, th...

Consumer preferences, and potential health risks associated with the consumption of synthetic food colors, explain the commercial interest in alternatives from natural sources. Betaxanthins from cactus fruit have been used to color food products, but sugars are not usually removed from these extracts leading to processing and formulation challenges...

Monoliths are considered the fourth generation of chromatographic supports, and they consist of a single piece of highly interconnected network of channels, distinguishing them from other chromatographic supports. The advantages of these supports have been exploited to separate or immobilize biomolecules including antibodies. Antibodies are highly...

PEGylated or polyethylene glycol-modified proteins have been used as therapeutic agents in different diseases. However, the major drawback in their procurement is the purification process to separate unreacted proteins and the PEGylated species. Several efforts have been done to separate PEGylation reactions by chromatography using different statio...

BACKGROUND A short processing time and efficient scale‐up stem cell isolation bioprocess is essential to exploit the potential of these cells for the treatment of multiple chronic diseases. Various methodologies have been used for stem cell recovery. However, most of them present economic and/or time‐consuming drawbacks. RESULTS This paper reports...

Cell‐based aqueous two‐phase system technologies present an emerging and promising translational tool for the development of next‐generation therapeutic applications. Advantages such as biocompatibility, scalability and ease of implementation confirm this possibility. The aim of this perspective article is to present a brief, process‐oriented analy...

Laccases are currently employed in several biotechnological applications such as pharmaceutical, environmental, textile and petrochemical. Their production in submerged bioreactors has been extensively reported, however, this approach exhibits a major drawback: the concomitant production of proteases, which reduce laccase activity in broths. In thi...

Poultry products are one of the major transmission media of Salmonella enteritidis to humans. A promising alternative to reduce the load of Salmonella in poultry are bacteriophages. Elsewhere, a mixture of six bacteriophages has been used successfully, but large‐scale production would be necessary to supply potential poultry market and costs analys...

In this study, the intensification of C-phycocyanin protein extraction from the green microalgae Spirulina maxima was successfully achieved using aqueous two-phase systems coupled to different continuous-operation extractors. Three systems with different total concentrations of Polyethylene glycol and potassium phosphate salt were tested in three d...

Neutropenia is a common side-effect of acute myeloid leukemia (AML) chemotherapy characterized by a critical drop in neutrophil blood concentration. Neutropenic patients are prone to infections, experience poorer clinical outcomes, and require expensive medical care. Although transfusions of donor neutrophils are a logical solution to neutropenia,...

Fractionation of native, mono and di-PEGylated lysozyme was performed in 36 different polymer-polymer aqueous two-phase systems using UCON as a phase-forming component. After a discrete partition analysis, dextran 75 kDa-UCON, volume ratio 3, tie-line length 35% w/w; ficoll 70 kDa-UCON, volume ratio 1, tie-line length 45% w/w and a PEG 8 kDa-UCON v...

Aqueous two-phase systems (ATPS) have been reported as an attractive biocompatible extraction system for recovery and purification of biological products. In this work, the implementation, characterization, and optimization (operational and economic) of invertase extraction from spent brewery yeast in a semi-automatized pilot plant using ATPS is re...

Contamination with Salmonella can cause significant economic and public health problems. Its prevention is accomplished by the use of antibiotics, but appearance of resistant strains has raised the need to search for alternatives. Recently, Salmonella-specific bacteriophages were identified and suggested as a therapy product in poultry. The large-s...

BACKGROUND Invertase use in food industry is limited by production costs. Alternative strategies for extraction, such as aqueous two‐phase systems (ATPS) and three‐phase partitioning (TPP), could be economically feasible for yeast invertase. Economic modeling of bioprocesses make possible the identification of critical parameters for production cos...

Aqueous two‐phase systems (ATPS) have proved to be an efficient and integrative operation to enhance recovery of industrially‐relevant bioproducts. After ATPS discovery, a variety of works have been published regarding their scaling from 10 to 1,000 liters. Although ATPS have achieved high recovery and purity yields, there is still a gap between th...

Aqueous two‐phase systems (ATPS) have been widely exploited for the recovery and partial purification of biological compounds. Recently our research group characterized the primary recovery and partial purification of bacteriophage M13 using polymer‐salt and ionic liquid‐salt ATPS. From such study, it was concluded that PEG 400‐potassium phosphate...

Antibiotics are a key pharmaceutical to inhibit growth or kill microorganisms. They represent a profitable market and, in particular, tetracycline has been listed as an essential medicine by the WHO. Therefore it is important to improve their production processes. Recently novel and traditional aqueous two-phase systems for the extraction have been...

Current chapter reviews the applications of ELISA in various different fields including food industry, vaccine development, immunology (autoimmunity and humoral immunity), diagnosis (pregnancy, cancer and infectious diseases), toxicology, drug monitoring, pharmaceutical industry, and transplantation. Different examples related to each area are expl...

This chapter presents the most commons errors that typically occur when performing ELISA. Following every error, the chapter points out the possible reasons behind such errors and possible methods to overcome the problems. The chapter also describes key important parameters in assay evaluation including sensitivity, specificity, accuracy and limit...

Current chapter describes the essential components of ELISA including the solid phase, the adsorbents (different types of target biomolecules), and the washing and blocking agents used in assay procedure. The chapter also reviews widely applied enzymes and substrates with their specific characteristics. To complete the assay, the chapter offers inf...

Nowadays ELISA is considered to be the troy horse for the routine clinical practice. This widely applied technique offers specific detection of a wide variety of target analytes in different kinds of samples. Since the invention of the technique four decades ago, ELISA has rapidly found various applications in food quality, environmental, biotechno...

Current chapter reviews background and history of the immunoassays until invention of ELISA. In that perspective, important evolutions in the field such as side-chain theory, antigen-antibody theory, discovery of the antibody structure, invention of radioimmunoassay (RIA), and invention of enzyme linked immunosorbent assay (ELISA) are reviewed. The...

BACKGROUND A laboratory‐scale protocol for the production, primary recovery and partial purification of fucoxanthin via biomass production at low light intensities was previously reported. This proposed approach exploited the use of ethanol salt aqueous two‐phase systems (ATPS) and ultrafiltration (UF) to deliver a 45% total recovery yield if all s...

Traditionally, ATPS have been used as a liquid-liquid extraction technique for the primary recovery and purification of biological samples. The enormous potential of their usage comes with great economical and technical advantages mainly due to the mild physicochemical environment. Nowadays, the use of ATPS as a bioengineering technique is approach...

Microalgae have been used during the past four decades in the Bio-industries for the production of high added value products and development of useful approaches with environmental applications. The fast growing rate, simple growth requirements and using sunlight as the major source of energy are the key factors for usage of algae. In the past 15 y...

Aqueous two-phase systems (ATPS) are regarded as an effective primary recovery and partial purification technique for biological products. However, although much has been developed through the last 50 years, there are still advances to address concerning the handling, improvement, and alternative uses of ATPS. Therefore, although ATPS has proven to...

Aqueous two-phase systems (ATPS) have been studied and used for product recovery and purification from diverse biological sources. ATPS are characterized by their versatility, easy scale-up parameters, process integration capability, and relative low cost. This technique is commonly regarded as a primary recovery stage mainly due to its low selecti...

Research has demonstrated how successful ATPS is to recover or purify different bioproducts, including proteins. They pose several advantages, which include cheap and nontoxic materials for their construction and linear scale-up, and it does not require a complex equipment for operation. All of these have the potential to help ATPS to decrease prod...

Aqueous two-phase systems (ATPS) are biphasic systems composed, traditionally, by two polymers or one polymer and one salt. However, the classification currently includes ATPS formed by ethanol, micelles, and ionic liquids. ATPS are a liquid–liquid separation technology used as primary recovery technique. This technology has been used to separate a...

Characterization of aqueous two-phase systems is an important procedure crucial for their successful operation. In these systems, the partition behavior of the solutes not only depends on their physicochemical properties but on the selected system design parameters which vary in a case-by-case basis. The selection of the phase-forming chemicals, th...

BACKGROUND Bacteriophage M13 is an Escherichia coli‐specific non‐lytic filamentous virus commonly used in applications ranging from antibody screening and nanomaterial construction to drug delivery, among others. In this tenor, alternative methods for the fractionation, recovery and partial purification of phage particles are desired. In this work,...

Protein refolding is a strategy used to obtain active forms of proteins from inclusion bodies. On its part, laccase is an enzyme with potential for different biotechnological applications but there are few reports regarding its refolding which in many cases is considered inefficient due to the poor obtained refolding yields. Aqueous Two-Phase Syste...

BACKGROUND Purification of PEGylated proteins from reactions is still a challenge due to the formation of isomers. In this work, the recovery of PEGylated lysozyme was studied in a novel integrative approach called in situ ATPS. The excess of PEG in the lysozyme PEGylation reaction (LPR) was used as part of the phase forming chemicals with differen...

BACKGROUND The bioavailability of therapeutic proteins is improved through PEGylation. This chemical modification involves the production of isomers with different numbers and sites of attached PEG chains, which are difficult to separate efficiently. Their purification with chromatography requires an understanding of the operation and the evaluatio...

The use of polygalacturonase (PG) preparations in winemaking promotes the release of phenolic compounds. A PG from a new source, Aspergillus sojae mutant, was semi-purified and tested for grape mash maceration. Crude extract (CE), a commercial pectinase, and two high PG activity semi-purified preparations, FI and FII, were applied for maceration at...

BACKGROUND The efficient, controlled and robust purification of conjugates from PEGylation has a growing demand in the biopharmaceutical's market. In general, the yield and purity reached through the conventional chromatographic modes are not particularly high or efficient. Affinity chromatography has so far scarcely been explored. The present work...

The cover image, by Marco Rito‐Palomares et al., is based on the Perspective Monolithic chromatography: insights and practical perspectives, DOI: 10.1002/jctb.5040.

This comprehensive and unique text presents a full overview of downstream processing useful for those new to the concept as well as professionals with experience in the area. The history and theoretical principles of Aqueous Two-Phase Systems (ATPS) are covered in depth. Information on ATPS characterization and application is included, and ATPS equ...

Aqueous two-phase systems (ATPS) have been developed as an intensive unit operations that can be easily scaled-up and doesn’t require complex equipment for operation for protein purification. Although ATPS has been used successfully to recover/purify diverse bioproducts, there is a lack of information on how they affect production costs. The presen...

B-Phycoerythrin is a color protein with several applications, from food coloring to molecular labeling. Depending on the application, different purity is required, affecting production cost and price. Different production and purification strategies for B-Phycoerythrin have been developed, the most studied are based on the production using Porphyri...

The use of monoliths, one-piece porous separation materials, as column packaging media is known as monolithic chromatography and this technology has been widely exploited to improve the separation profile of high-valued biomolecules from complex biological matrices. Monoliths possess a wide range of processing advantages over commonly used chromato...

Aqueous two-phase systems (ATPS) have emerged as an alternative strategy for the recovery and purification of a wide variety of biological products. Typical process development requires a large screening of experimental conditions towards industrial adoption where continuous processes are preferred. In this work, it was proved that under certain fl...

Abbreviations: ANOVA, analysis of variance; CE, coupling efficiency; IAC, immunoaffinity chromatography; NHS, n-hydroxysuccinimide.ABSTRACTImmunosorbents in Inmunoaffinity Chromatography are prepared immobilizing expensive antibodies without guidelines for ensuring the best coupling efficiencies, and avoid low binding capacities. Covalent immobiliz...

In this work, we demonstrate that signal-masking reagents together with appropriate capture antibody carriers can eliminate the washing steps in sandwich immunoassays. A flotation immunoassay (FI) platform was developed with horseradish peroxidase chemiluminescence as the reporter system, the dye Brilliant Blue FCF as the signal-masking reagent, an...

BACKGROUND Multiple aqueous two-phase systems (ATPS) were evaluated for the partial purification of laccase from a crude aqueous extract. The laccase extract was obtained from residual compost, generated from the production of the edible mushroom Pleurotus ostreatus. A total of 59 ATPS were evaluated; 16 systems using poly (ethylene glycol) (PEG) a...

Protein hydrophobicity can be modified after a PEGylation process. However, hydrophobic interaction chromatography (HIC) has been used to separate PEGylation reaction products less frequently than other techniques. In this context, chromatographic monoliths represent a good alternative to continue exploring the separation of PEGylated proteins with...

Aqueous two-phase systems (ATPS) are a liquid-liquid extraction technology with clear process benefits; however, its lack of industrial embracement is still a challenge to overcome. Antibodies are a potential product to be recovered by ATPS in a commercial context. The objective of this work is to present a more integral approach of the different i...

The increasing attention to biological activities of dietary xanthophylls demands the establishment of attractive and easy-to-implement purification protocols for their recovery from different sources. Bioengineering strategies that involve alternative isolation and purification techniques have gained interest since they offer practical benefits in...

Contamination by cell debris and non-viable cells is common in cell-based processes. Cell fragments and dead cells can be produced during culture and in purification steps, and often must be eliminated before analyses, subsequent process steps, and/or final application to avoid interference, fouling and reduced product yields. In the present work,...

Uricase is the enzyme responsible for the breakdown of uric acid, the key molecule leading to gout in humans, into allantoin, but it is absent in humans. It has been produced as a PEGylated pharmaceutical where the purification is performed through three sequential chromatographic columns. More recently an aqueous two-phase system (ATPS) was report...