Field Of Biotechnology Research Articles

Free-Volume analysis of PAMAM dendrimers by means of positron annihilation lifetime spectroscopy

Polymeric structures known as dendrimers branch around a nucleus and contain functional groups that can affix surface groups to their ends. The inherent structural free volume of these materials makes them attractive for various important applications in the fields of biotechnology and nanotechnology. The purpose of this research was to examine the free volume behavior of ester- and acid-terminated poly(amidoamine) (PAMAM) dendrimers cored with tris(2-aminoethyl)amine (TREN) for the first three generations (0.5, 1.5, and 2.5 generations). Positron annihilation life spectroscopy (PALS) was used to analyze and discuss the size and fractions of hole-free volume generated by thermal effects in the temperature range of 10 °C to 80 °C. In addition, these PALS results may be related to the entanglement and deformability of the generation-dependent architecture and the influence of the surface groups.

Quenching Reaction-Diffusion Systems in Bioengineering and Life Sciences

This research paper revolves around investigating the phenomenon of quenching in reaction-diffusion systems and highlighting its significance. The primary focus is on analyzing a specific type of parabolic singular reaction-diffusion model that incorporates positive Dirichlet boundary conditions. The objective is to establish the sufficiency of certain conditions for quenching to occur within a finite time frame and to demonstrate the global existence of solutions. The novelty of this paper lies in the simplicity of the conditions imposed on the nonlinearity. This simplicity allows us to choose it from a wide range of possibilities, thus facilitating the application of the model to numerous singular reaction-diffusion phenomena. To bolster our findings, we will present various real-world applications in the fields of bioengineering and life sciences, showcasing the practical relevance of quenching phenomena. Finally, the paper ends with a conclusion and some potential future perspectives for further research in this area.

Bioprospection of the Antarctic Diatoms Craspedostauros ineffabilis IMA082A and Craspedostauros zucchelli IMA088A.

In extreme environments such as Antarctica, a diverse range of organisms, including diatoms, serve as essential reservoirs of distinctive bioactive compounds with significant implications in pharmaceutical, cosmeceutical, nutraceutical, and biotechnological fields. This is the case of the new species Craspedostauros ineffabilis IMA082A and Craspedostauros zucchellii IMA088A Trentin, Moschin, Lopes, Custódio and Moro (Bacillariophyta) that are here explored for the first time for possible biotechnological applications. For this purpose, a bioprospection approach was applied by preparing organic extracts (acetone and methanol) from freeze-dried biomass followed by the evaluation of their in vitro antioxidant properties and inhibitory activities on enzymes related with Alzheimer's disease (acetylcholinesterase: AChE, butyrylcholinesterase: BChE), Type 2 diabetes mellitus (T2DM, α-glucosidase, α-amylase), obesity (lipase) and hyperpigmentation (tyrosinase). Extracts were then profiled by ultra-high-performance liquid chromatography-mass spectrometry (UPLC-HR-MS/MS), while the fatty acid methyl ester (FAME) profiles were established by gas chromatography-mass spectrometry (GC-MS). Our results highlighted strong copper chelating activity of the acetone extract from C. ineffabilis and moderate to high inhibitory activities on AChE, BChE, α-amylase and lipase for extracts from both species. The results of the chemical analysis indicated polyunsaturated fatty acids (PUFA) and their derivatives as the possible compounds responsible for the observed activities. The FAME profile showed saturated fatty acids (SFA) as the main group and methyl palmitoleate (C16:1) as the predominant FAME in both species. Overall, our results suggest both Antarctic strains as potential sources of interesting molecules with industrial applications. Further studies aiming to investigate unidentified metabolites and to maximize growth yield and natural compound production are required.

Protein Cryoprotectant Ability of the Aqueous Zwitterionic Solution.

Protein cryopreservation is important for the long-term storage of unstable proteins. Recently, we found that N-acetylglucosaminyltransferase-V (GnT-V) can be cryopreserved in a deep freezer without temperature control using a dilute binary aqueous solution of 3-(1-(2-(2-methoxyethoxy)ethyl)imidazol-3-io)butane-1-carboxylate (OE2imC3C) [10 wt %, mole fraction of solute (x) = 7.75 × 10-3], an artificial zwitterion. However, it is unclear which solvent properties are required in these media to preserve unstable proteins, such as GnT-V. In this study, we investigated the melting phenomena and solution structure of dilute binary aqueous OE2imC3C solutions [x = 0-2.96 × 10-2 (0-30 wt %)] using differential scanning calorimetry (DSC) and Raman and Fourier transform infrared (FTIR) spectroscopies combined with molecular dynamics (MD) simulation to compare the cryoprotectant ability of OE2imC3C with two general cryoprotectants (CPAs), glycerol and dimethyl sulfoxide. DSC results indicated that aqueous OE2imC3C solutions can be melted at lower temperatures with less energy than the control CPA solution, with increasing x, primarily due to OE2imC3C having a higher content of unfrozen water molecules. Moreover, Raman and FTIR results showed that the high content of unfrozen water molecules in aqueous OE2imC3C solutions was due to the hydration around the ionic parts (the COO- group and imidazolium ring) and the OCH2CH2O segment. In addition, the MD simulation results showed that there were fewer structured water molecules around the OCH2CH2O segment than the hydration water molecules around the ionic parts. These solvent properties suggest that dilute aqueous OE2imC3C solutions are effective in preventing freezing, even in a deep freezer. Therefore, this medium has the potential to act as a novel cryoprotectant for proteins in biotechnology and biomedical fields.

Complete microbial synthesis of crocetin and crocins from glycerol in Escherichia coli

BackgroundCrocin, a glycosylated apocarotenoid pigment predominantly found in saffron, has garnered significant interest in the field of biotechnology for its bioactive properties. Traditional production of crocins and their aglycone, crocetin, typically involves extraction from crocin-producing plants. This study aimed to develop an alternative biosynthetic method for these compounds by engineering the metabolic pathways of zeaxanthin, crocetin, and crocin in Escherichia coli strains.ResultsEmploying a series of genetic modifications and the strategic overexpression of key enzymes, we successfully established a complete microbial pathway for synthesizing crocetin and four glycosylated derivatives of crocetin, utilizing glycerol as the primary carbon source. The overexpression of zeaxanthin cleavage dioxygenase and a novel variant of crocetin dialdehyde dehydrogenase resulted in a notable yield of crocetin (34.77 ± 1.03 mg/L). Further optimization involved the overexpression of new types of crocetin and crocin-2 glycosyltransferases, facilitating the production of crocin-1 (6.29 ± 0.19 mg/L), crocin-2 (5.29 ± 0.24 mg/L), crocin-3 (1.48 ± 0.10 mg/L), and crocin-4 (2.72 ± 0.13 mg/L).ConclusionsThis investigation introduces a pioneering and integrated microbial synthesis method for generating crocin and its derivatives, employing glycerol as a sustainable carbon feedstock. The substantial yields achieved highlight the commercial potential of microbial-derived crocins as an eco-friendly alternative to plant extraction methods. The development of these microbial processes not only broadens the scope for crocin production but also suggests significant implications for the exploitation of bioengineered compounds in pharmaceutical and food industries.

Achievement of India in the field of Bio-Technology

India has made remarkable achievements in the field of biotechnology, spanning healthcare, agriculture, industrial bioprocessing, and environmental sustainability. In healthcare, the country has emerged as a global leader in producing affordable biopharmaceuticals, vaccines, and diagnostic tools, addressing critical healthcare challenges. In agriculture, India’s expertise in genetically modified crops, crop improvement, and seed technology has revolutionized farming practices, enhancing productivity and sustainability. Industrial biotechnology has witnessed significant growth, with Indian companies excelling in enzyme production, biofuels, and waste management solutions. Additionally, India has made notable strides in environmental biotechnology, employing innovative methods for wastewater treatment, biogas production, and bioremediation. Despite challenges, India’s biotechnology sector is poised for further growth, driven by advancements in emerging technologies and collaborative efforts among stakeholders. These achievements underscore India’s potential to contribute significantly to global biotechnological innovation and address pressing societal and environmental challenges. Biotechnology refers to all those techniques which use biological processes to produce substances which can be used as medicines and in industries. It also includes all those methods (and their study) by which a species of living beings (especially human species) can be made superior. For example, making vaccines to prevent diseases, making/using bio-fertilizers, making/using biopesticides, making superior species of plants through bio-engineering, protecting the environment and biodiversity, development of DNA figure printing technology, cloning etc. all come under biotechnology. Modern biotechnological processes have their origins in the techniques used in ancient times, such as making vinegar byfermenting sugarcane juice, obtaining curd by coagulating milk, making cheese, etc. Similarly, in the field of agriculture, new varieties of seeds are prepared by hybridizing different types of plants, etc.

Generating Retinas through Guided Pluripotent Stem Cell Differentiation and Direct Somatic Cell Reprogramming.

Retinal degeneration diseases affect millions of people worldwide but are among the most difficult eye diseases to cure. Studying the mechanisms and developing new therapies for these blinding diseases requires researchers to have access to many retinal cells. In recent years there has been substantial advances in the field of biotechnology in generating retinal cells and even tissues in vitro, either through programmed sequential stem cell differentiation or direct somatic cell lineage reprogramming. The resemblance of these in vitro-generated retinal cells to native cells has been increasingly utilized by researchers. With the help of these in vitro retinal models, we now have a better understanding of human retinas and retinal diseases. Furthermore, these in vitro-generated retinal cells can be used as donor cells which solves a major hurdle in the development of cell replacement therapy for retinal degeneration diseases, while providing a promising option for patients suffering from these diseases. In this review, we summarize the development of pluripotent stem cell-to-retinal cell differentiation methods, the recent advances in generating retinal cells through direct somatic cell reprogramming, and the translational applications of retinal cells generated in vitro. Finally, we discuss the limitations of the current protocols and possible future directions for improvement.

Exploring the Function of Gene Promoter Regulatory Elements Using CRISPR Tools.

Gene promoters serve as pivotal regulators of transcription, orchestrating the initiation, rate, and specificity of gene expression, resulting in cellular diversity found among distinct cell types within multicellular organisms. Identification of the sequence and function of promoters' regulatory elements and their complex interaction with transcription factors, enhancers, silencers, and insulators is fundamental to coordinated transcriptional processes within cells. Identifying these regulatory elements and scrutinizing their functions and interactions through the use of synthetic promoters can pave the way for researchers in various fields ranging from uncovering the origins of diseases associated with promoter mutations to harnessing these regulatory components in biotechnological applications.In this chapter, we describe the manipulation of regulatory elements within promoters, with a specific focus on the use of CRISPR technology on enhancers and silencer elements of the Ovalbumin gene promoter. We explain and discuss processes for the deletion of/interference with regulatory elements within the promoter, employing CRISPR-based approaches. Furthermore, we demonstrate that a CRISPR/Cas-manipulated promoter can activate gene transcription in cell types where it is normally inactive. This confirms that CRISPR technology can be effectively used to engineer synthetic promoters with desired characteristics, such as inducibility, tissue-specificity, or enhanced transcriptional strength. Such an approach provides valuable insights into the mechanisms and dynamics of gene expression, thereby offering new opportunities in the fields of biotechnology and medicine.

Self-indicating polymers: a pathway to intelligent materials.

Self-indicating polymers have emerged as a promising class of smart materials that possess the unique ability to undergo detectable variations in their physical or chemical properties in response to various stimuli. This article presents an overview of the most important mechanisms through which these materials exhibit self-indication, including aggregation, phase transition, covalent and non-covalent bond cleavage, isomerization, charge transfer, and energy transfer. Aggregation is a prevalent mechanism observed in self-indicating polymers, where changes in the degree of molecular organization result in variations in optical or electrical properties. Phase transition-induced self-indication relies on the transformation between different phases, such as liquid-to-solid or crystalline-to-amorphous transitions, leading to observable changes in color or conductivity. Covalent bond cleavage-based self-indicating polymers undergo controlled degradation or fragmentation upon exposure to specific triggers, resulting in noticeable variations in their structural or mechanical properties. Isomerization is another crucial mechanism exploited in self-indicating polymers, where the reversible transformation between the different isomeric forms induces detectable changes in fluorescence or absorption spectra. Charge transfer-based self-indicating polymers rely on the modulation of electron or hole transfer within the polymer backbone, manifesting as changes in electrical conductivity or redox properties. Energy transfer is an essential mechanism utilized by certain self-indicating polymers, where energy transfer between chromophores or fluorophores leads to variations in the emission characteristics. Furthermore, this review article highlights the diverse range of applications for self-indicating polymers. These materials find particular use in sensing and monitoring applications, where their responsive nature enables them to act as sensors for specific analytes, environmental parameters, or mechanical stress. Self-indicating polymers have also been used in the development of smart materials, including stimuli-responsive coatings, drug delivery systems, food sensors, wearable devices, and molecular switches. The unique combination of tunable properties and responsiveness makes self-indicating polymers highly promising for future advancements in the fields of biotechnology, materials science, and electronics.

Assessment of effectiveness of practice teaching biology methods in collective learning: The pilot study

This study constitutes a pilot investigation conducted in the absence of a control group. Its objective was to evaluate the efficacy of practical learning within a collaborative setting among first-year students specializing in biotechnology. The research session focused on the topic “Methods for the development and production of biological products: The role of biological products in human life.” Sixteen students were divided into four groups of four students each. The lesson was facilitated using the Jigsaw group teaching method, integrating both theoretical and practical components. Pre- and post-class tests were administered to evaluate students' performance, revealing significant improvements after the session.The assessment of students who underwent the post-class test significantly exceeded their performance in the pre-class test. For theoretical questions, this yielded: T=14.478, P<0.001, and for practical questions: T=19.104, P<0.001. Additionally, the study revealed that the implementation of the Jigsaw method leads to a substantial increase in the percentage of students achieving high grades and a significant decrease in the proportion of students receiving low grades. The percentages of students receiving high grades increased by 43.75% (excellent) and 6.25% (good), while the percentage of low grades decreased by 50% (fair) and 12.5% (bad). This study provides empirical evidence supporting the utilization of collaborative, experiential learning approaches in enhancing biology education, particularly in fostering both theoretical understanding *and practical skills among students in the field of biotechnology.

Engineering high affinity antigen-binders: Beyond conventional antibodies.

For decades, antibodies have remained the archetypal binding proteins that can be rapidly produced with high affinity and specificity against virtually any target. A conventional antibody is still considered the prototype of a binding molecule. It is therefore not surprising that antibodies are routinely used in basic scientific and biomedical research, analytical workflows, molecular diagnostics etc. and represent the fastest growing sector in the field of biotechnology. However, several limitations associated with conventional antibodies, including stringent requirement of animal immunizations, mammalian cells for expression, issues on stability and aggregation, bulkier size and the overall time and cost of production has propelled evolution of concepts along alternative antigen binders. Rapidly evolving protein engineering approaches and high throughput screening platforms have further complemented the development of myriads of classes of non-conventional protein binders including antibody derived as well as non-antibody based molecular scaffolds. These non-canonical binders are finding use across disciplines of which diagnostics and therapeutics are the most noteworthy.

Microalgae as a potential natural source for the green synthesis of nanoparticles.

The increasing global population is driving the development of alternative sources of food and energy, as well as better or new alternatives for health and environmental care, which represent key challenges in the field of biotechnology. Microalgae represent a very important source material to produce several high-value-added bioproducts. Due to the rapid changes in the modern world, there is a need to build new materials for use, including those in the nanometer size, although these developments may be chronological but often do not occur at a time. In the last few years, a new frontier has opened up at the interface of biotechnology and nanotechnology. This new frontier could help microalgae-based nanomaterials to possess new functions and abilities. Processes for the green synthesis of nanomaterials are being investigated, and the availability of biological resources such as microalgae is continuously being examined. The present review provides a concise overview of the recent advances in the synthesis, characterization, and applications of nanoparticles formed using a wide range of microalgae-based biosynthesis processes. Highlighting their innovative and sustainable potential in current research, our study contributes towards the in-depth understanding and provides latest updates on the alternatives offered by microalgae in the synthesis of nanomaterials.

Analysis of total quality management (TQM) implementation in the biotechnology industry

Over the past two decades, the relevance of the Total Quality Management (TQM) model has increased significantly. TQM is becoming increasingly popular as organizations focus more on improving the quality of their products, services, and internal operations to increase customer value and gain competitive advantage. Organizations from the biotechnology sector play a significant role in the development of the national economy, the social sphere and employment. In such a situation, they face strong competitive advantages of competitors from all over the world, such as low costs and high productivity. The change in the amount of added value alone over the last 10 years leads to an increase of 35.2% per year. In 2009 is BGN 12 million, and in 2019 is BGN 208 million. That is why it is critical for them to improve their organizational performance through the Total Quality Management (TQM) approach. The purpose of this paper is to analyze the individual components of TQM in organizations from the field of biotechnology and food industry and to determine its model in terms of engineering activities. Pre-designed questionnaires were used as a data collection instrument in the present study. Conducting semi-structured interviews with team leaders in a biotech manufacturing organization provides insight into the impact of TQM on competitiveness. These elements, leadership skills and supplier/customer relationships have been found to play an important role in improving overall performance.

Enhancing small-scale acetification processes using adsorbed Acetobacter pasteurianus UMCC 2951 on κ-carrageenan-coated luffa sponge.

This study explored the utilization of luffa sponge (LS) in enhancing acetification processes. LS is known for having high porosity and specific surface area, and can provide a novel means of supporting the growth of acetic acid bacteria (AAB) to improve biomass yield and acetification rate, and thereby promote more efficient and sustainable vinegar production. Moreover, the promising potential of LS and luffa sponge coated with κ-carrageenan (LSK) means they may represent effective alternatives for the co-production of industrially valuable bioproducts, for example bacterial cellulose (BC) and acetic acid. LS and LSK were employed as adsorbents for Acetobacter pasteurianus UMCC 2951 in a submerged semi-continuous acetification process. Experiments were conducted under reciprocal shaking at 1 Hz and a temperature of 32°C. The performance of the two systems (LS-AAB and LSK-AAB respectively) was evaluated based on cell dry weight (CDW), acetification rate, and BC biofilm formation. The use of LS significantly increased the biomass yield during acetification, achieving a CDW of 3.34 mg/L versus the 0.91 mg/L obtained with planktonic cells. Coating LS with κ-carrageenan further enhanced yield, with a CDW of 4.45 mg/L. Acetification rates were also higher in the LSK-AAB system, reaching 3.33±0.05 g/Ld as opposed to 2.45±0.05 g/Ld for LS-AAB and 1.13±0.05g/Ld for planktonic cells. Additionally, BC biofilm formation during the second operational cycle was more pronounced in the LSK-AAB system (37.0±3.0 mg/L, as opposed to 25.0±2.0 mg/L in LS-AAB). This study demonstrates that LS significantly improves the efficiency of the acetification process, particularly when enhanced with κ-carrageenan. The increased biomass yield, accelerated acetification, and enhanced BC biofilm formation highlight the potential of the LS-AAB system, and especially the LSK-AAB variant, in sustainable and effective vinegar production. These systems offer a promising approach for small-scale, semi-continuous acetification processes that aligns with eco-friendly practices and caters to specialized market needs. Finally, this innovative method facilitates the dual production of acetic acid and bacterial cellulose, with potential applications in biotechnological fields.

Aqueous two-phase systems - versatile and advanced (bio)process engineering tools.

Aqueous two-phase systems (ATPS), also known as Aqueous Biphasic Systems (ABS), have been extensively studied as platforms for the separation and purification of biomolecules and other valuable compounds. These liquid-liquid extraction (LLE) systems have been a tool for biotechnology since its origin (Albertsson, 1950's), recently expanding to exciting fields such as health, biomedicine and material sciences. Due to their biocompatibility, amenability, flexibility, and versatility, ATPS have been applied across various research areas, addressing many challenges associated with conventional methodologies. In this feature article, we first discuss the fundamentals of ATPS and the molecular mechanisms that govern their formation and are crucial for their application. We then explore the most prominent and innovative applications of these systems in downstream processing. Additionally, we provide insights into the design of in situ upstream-downstream integrated platforms, and their use as pre-treatment and analytical tools. The latest advancements in ATPS applications within disruptive bioengineering and biotechnology fields are presented, along with their pioneering use in emerging scientific areas, such as the formation of all-aqueous (water-in-water) emulsions, microfluidic systems, and membrane-free batteries. Overall, this work underscores the transformative potential of ATPS in various branches of science, pinpointing directions for future research to fully explore and maximize ATPS capabilities, overcome existing hurdles, and drive innovation forward.

Towards biotechnological production of bio-based low molecular weight esters: a patent review.

Low molecular weight (LMW) esters, like ethyl acetate, methyl formate or butyl acetate, are widespread bulk chemicals in many industries. Each of them is currently produced in huge amounts (millions of tons per year scale) starting from fossil-based feedstock and they are used mainly because of their low toxicity and complete biodegradability. Energy transition is just half of the story on the path of fighting climate change: 45% of the global greenhouse gas emissions are caused by the production and use of all the products, materials and food necessary for modern human life. If the world is to reach its climate goals, there is the need to leave underground a significant proportion of the fossil feedstock and minimize environmental impacts of chemical manufacturing. This is the reason why a lot of efforts have been made to find novel routes for LMW esters production starting from renewable raw materials (e.g. biomasses or off-gases) and exploiting low-impact manufacturing, such as microbial fermentation or enzymatic reactions. This review reports the most significant patents, in the field of white biotechnology, that will hopefully lead to the commercialization of bio-based LMW esters as well as novel strategies, current problems to be solved, newer technologies, and some patent applications aiming at possible future developments.

Sustainable microalgae cultivation: A comprehensive review of open and enclosed systems for biofuel and high value compound production

Microalgae have emerged as a promising feedstock for biofuels and high-value compounds, offering potential solutions to global energy and resource challenges. This comprehensive review examines the latest advancements in microalgae cultivation technologies, focusing on both open systems and enclosed photobioreactors (PBRs). We analyze various configurations including open raceway ponds, tubular PBRs, flat panel PBRs, and novel designs such as the Light Exchange Bubble-column (LEB). The review encompasses key performance indicators such as biomass productivity, energy efficiency, and water usage, as well as life cycle assessment (LCA) results for different cultivation systems. We also discuss the challenges and opportunities in scaling up microalgae production, the potential for integrating wastewater treatment and CO2 mitigation, and the prospects of biorefinery approaches. By synthesizing recent research findings and identifying knowledge gaps, this review aims to provide a comprehensive understanding of the current state and future directions in sustainable microalgae cultivation for researchers, engineers, and policymakers in the field of renewable energy and biotechnology.

Applications, advancements, and challenges of 3D bioprinting in organ transplantation.

To date, organ transplantation remains an effective method for treating end-stage diseases of various organs. In recent years, despite the continuous development of organ transplantation technology, a variety of problems restricting its progress have emerged one after another, and the shortage of donors is at the top of the list. Bioprinting is a very useful tool that has huge application potential in many fields of life science and biotechnology, among which its use in medicine occupies a large area. With the development of bioprinting, advances in medicine have focused on printing cells and tissues for tissue regeneration and reconstruction of viable human organs, such as the heart, kidneys, and bones. In recent years, with the development of organ transplantation, three-dimensional (3D) bioprinting has played an increasingly important role in this field, giving rise to many unsolved problems, including a shortage of organ donors. This review respectively introduces the development of 3D bioprinting as well as its working principles and main applications in the medical field, especially in the applications, and advancements and challenges of 3D bioprinting in organ transplantation. With the continuous update and progress of printing technology and its deeper integration with the medical field, many obstacles will have new solutions, including tissue repair and regeneration, organ reconstruction, etc., especially in the field of organ transplantation. 3D printing technology will provide a better solution to the problem of donor shortage.

Research and Recent Achievements in Agriculture and Biotechnology with Innovative Technologies Application

This paper brings together researchers, scientists and industry experts to present the latest achievements in the dynamically developing fields of agriculture and biotechnology. Special attention was paid to interdisciplinary cooperation, advanced technologies and data-based solutions aimed at solving problems of food security, environmental sustainability and agricultural efficiency. Key topics included precision farming, gene editing, advanced breeding techniques, and the transformative role of big data analytics, artificial intelligence, and machine learning in optimizing agricultural processes. In light of this, to address numerous challenges in agricultural culture, I have proposed and developed the idea of creating a game. The game “World of Agricultural Culture” will present users with a bright HD graphic design and exciting gameplay. The professional version of the simulator is focused on business management and development in the agricultural industry.This is an additional way to improve the infrastructure of the Russian Federation, despite the presence of international sanctions. Thus, the competitiveness and attractiveness of the state increases, contributing to the stable development of Russian agriculture and the elimination of negative risks in the foreign economic sphere of the agricultural sector. In the course of the conducted research, a high level of efficiency improvement in the agro-industrial sector of the economy was noted.

Microfluidic Cartridge for Bead-Based Affinity Assays.

Within the vast field of medical biotechnology, the biopharmaceutical industry is particularly fast-growing and highly competitive, so reducing time and costs associated to process optimization becomes instrumental to ensure speed to market and, consequently, profitability. The manufacturing of biopharmaceutical products, namely, monoclonal antibodies (mAbs), relies mostly on mammalian cell culture processes, which are highly dynamic and, consequently, difficult to optimize. In this context, there is currently an unmet need of analytical methods that can be integrated at-line in a bioreactor, for systematic monitoring and quantification of key metabolites and proteins. Microfluidic-based assays have been extensively and successfully applied in the field of molecular diagnostics; however, this technology remains largely unexplored for Process Analytical Technology (PAT), despite holding great potential for the at-line measurement of different analytes in bioreactor processes, combining low reagent/molecule consumption with assay sensitivity and rapid turnaround times.Here, the fabrication and handling of a microfluidic cartridge for protein quantification using bead-based affinity assays is described. The device allows geometrical multiplexed immunodetection of specific protein analytes directly from bioreactor samples within 2.5h and minimal hands-on time. As a proof-of-concept, quantification of Chinese hamster ovary (CHO) host cell proteins (HCP) as key impurities, IgG as product of interest, and lactate dehydrogenase (LDH) as cell viability marker was demonstrated with limits of detection (LoD) in the low ng/mL range. Negligible matrix interference and no cross-reactivity between the different immunoassays on chip were found. The results highlight the potential of the miniaturized analytical method for PAT at reduced cost and complexity in comparison with sophisticated instruments that are currently the state-of-the-art in this context.