Quality By Design Principles Research Articles (Page 1)

Biopharmaceutical manufacturing processes in which the product of interest is extracellularly expressed typically employ a clarification step following cell culture or fermentation. During clarification, crude cell culture fluid or fermentation broth is processed to remove insoluble solids, cells, debris, and other particulates, with the extracellular product of interest retained in the filtrate. Soluble impurities, such as host cell proteins (HCPs), may also be partially removed. Historically, the clarification process has been considered a limited contributor to Critical Quality Attributes (CQA). As part of upstream harvest, many biopharmaceutical companies have not fully developed quality control strategies from process development to manufacturing, complicating the application of Quality by Design (QbD) principles to this step. However, advancements in upstream and downstream processing (DSP) technologies, alongside increasing cell counts and titers, necessitate reevaluating clarification as a critical process contributing to drug product quality. Conducting controlled studies to define the process and establish parameters using QbD principles can improve control over process impurities and facilitate a logical quality control strategy, integrating quality into the process. This article describes a systematic approach to QbD for a harvest clarification process where the product of interest is extracellular and impurities are removed in the filtrate post-clarification. It highlights methods for optimizing the clarification unit operation using QbD principles, ensuring better process efficiency, and product quality.

Quality by Design (QbD) has emerged as a systematic and proactive approach in pharmaceutical development, ensuring consistent product quality through a thorough understanding of formulation components and critical process parameters. In the context of phytosomal formulations, which increase bioavailability and therapeutic effectiveness of phytoconstituents. QbD-driven risk analysis is essential for optimizing formulation parameters and reducing variability. The incorporation of risk assessment tools, such as Fault Tree Analysis (FTA), Ishikawa fishbone diagrams, Failure Mode and Effect Analysis (FMEA), and Design of Experiments (DoE), facilitates the identification and management of critical material attributes (CMAs) and critical process parameters (CPPs) that profoundly affect the quality attributes of phytosomal carriers. Utilizing a scientific and data-driven methodology, QbD enhances formulation development, resulting in superior stability, encapsulation efficiency, and controlled release properties. Furthermore, the utilization of QbD principles ensures regulatory adherence, improves repeatability, and minimizes batch-to-batch variability, resulting in a more dependable and scalable production process. The pharmaceutical industry is shifting to a methodical and knowledge-based approach, and QbD-driven risk analysis in phytosomal formulations is a transformational tool for maximizing the therapeutic potential of bioactive phytoconstituents.

Azithromycin (AZT), a macrolide antibiotic, has recently been explored as an injection therapy for osteoarthritis. However, its instability and poor solubility limit its effect due to an insufficient quantity and duration at the target sites. To address these challenges, this study developed poly (lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) for AZT delivery, which were subsequently incorporated into a thermoresponsive injectable hydrogel suitable for intra-articular administration. The formulation was developed using a Quality by Design (QbD) approach, focusing on two steps: (i) preparation of AZT-PLGA NPs and (ii) loading the NPs into a poloxamer-based hydrogel. Critical material attributes (AZT, PLGA, surfactants) were evaluated for their impacts on the critical quality attributes (CQAs) of the NP formulation (size distribution and encapsulation efficiency). The optimized AZT-PLGA NPs exhibited a mean particle size of ~ 150 nm and a PDI of < 0.2, ensuring uniformity and stability. Secondly, these NPs were then embedded into a novel thermoresponsive hydrogel. The effects of NPs, hyaluronic acid, and mannitol on physical appearance, thermal sensitivity, the rheology (shear-thinning and thixotropic), pH, and sustained release properties of the final formulation were systematically investigated. Fourier-transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) analyses revealed interactions between AZT and PLGA, which, while not affecting the drug assay, enhanced the structural integrity and modified the thermal properties of the final product. Using QbD principles, a risk-based assessment was proposed for future drug development. This study introduced a novel thermoresponsive injectable hydrogel for the intra-articular delivery of AZT using PLGA nanoparticles.

INTRODUCTION. Quality by design (QbD) is a systematic approach to pharmaceu­tical development that begins with predefined objectives and emphasises product and process understanding and process control, based on sound science and quality risk management. The QbD approach facilitates the production of medicinal products with target characteristics and quality profiles. There are currently no specific guidelines for the application of QbD principles to the development of individual dosage forms.AIM. This study aimed to evaluate the possibility of and propose an algorithm for using QbD at the laboratory stage of pharmaceutical development for solid dosage forms, with tablets as a case study.MATERIALS AND METHODS. This study analysed publicly available regulatory documents, scientific publications, and guidelines on pharmaceutical development using general scientific methods, including comparative and logical analysis. The regula­tory documents analysed included those issued by the International Council for Harmonisation (ICH), the Eurasian Economic Commission, and the State Pharmacopoeia of the Russian Federation. The sources searched included electronic databases, such as PubMed, Web of Science, eLIBRARY.RU, and Google Scholar.RESULTS. Developing the quality target product profile (QTPP) and composition of tablets requires a comprehensive study of the active substance, as well as an assessment of its compatibility with the excipients. At the laboratory stage of pharmaceu­tical development, it is necessary to select and optimise the medicinal product composition while assessing potential risks. This approach provides for the preliminary identification of critical quality attributes, critical process parameters, and critical material parameters. This article presents an algorithm for applying QbD to tablet formulations at the laboratory stage of pharmaceutical development.CONCLUSIONS. When implemented at the laboratory stage, the proposed algorithm with QbD elements will improve the overall efficiency of pharmaceutical development.

Background: Polymeric nanoparticles (PNPs) have emerged as promising drug delivery systems to overcome the limitations of conventional chemotherapeutics. Capecitabine, a prodrug of 5-fluorouracil (5-FU), is widely used in cancer therapy but suffers from poor bioavailability and systemic toxicity. The appli-cation of the Quality by Design (QbD) framework in PNP development provides a structured approach to address these challenges. Objective: This review examines the QbD principles in the formulation and optimization of capecitabine-loaded PNPs, focusing on strategies to enhance therapeutic efficacy and minimize adverse effects. Methods: The QbD approach encompasses defining a Quality Target Product Profile (QTPP), identifying Critical Quality Attributes (CQAs), and conducting risk assessments to pinpoint Critical Material Attributes (CMAs) and Critical Process Parameters (CPPs). Techniques such as Design of Experiments (DoE) facilitate systematic optimization. Results: Incorporating QbD principles ensures the development of robust PNP formulations with improved encapsulation efficiency, controlled drug release, and targeted delivery. Studies highlight the use of biodegradable polymers like PLGA, chitosan, and PEG for superior biocompatibility and stability. Analytical methods validate the consistency and quality of the nanoparticles. Conclusion: The QbD framework enables the rational design of capecitabine-loaded PNPs with enhanced bioavailability and reduced toxicity, contributing to safer and more effective cancer treatments. Future research should explore novel polymeric systems and advanced manufacturing technologies to expand the therapeutic potential of PNPs in oncology.

Quality by Design (QbD) is a structured approach to pharmaceutical development that ensures predefined product quality by understanding and controlling manufacturing processes from the outset. Unlike traditional methods focusing on end-product testing, QbD emphasizes building quality into the product design itself, enhancing manufacturing efficiency and regulatory compliance. This review highlights the application of QbD in developing generic solid oral drug products, emphasizing tools like risk assessment, process design, and control strategies to achieve consistent quality. Key components include identifying and managing Critical Quality Attributes (CQAs), Critical Process Parameters (CPPs), and Critical Material Attributes (CMAs), which influence formulation, process development, and overall performance. The integration of Design of Experiments (DoE) to systematically study the effects of multiple variables on product and process performance, enabling optimization and robust development and effective control strategies are also discussed in this article. Addressing regulatory expectations, particularly those from the International Council for Harmonisation (ICH), this review outlines how QbD principles help generics meet bioequivalence standards, ensuring consistent quality and performance. Applying QbD not only enhances product robustness and manufacturing efficiency but also improves patient safety through better process understanding and continuous improvement. This review article outlines the various steps involved in the development of generic drug products using the QbD approach from analysis of brand product to product lifecycle management and continual improvement.

Eberconazole nanostructured lipid carrier (EBR-NLC) 1% w/w optimization was done using the Quality by Design (QbD) approach, employing a 23 Full Factorial Design (FFD) for experimental planning, followed by thorough physico-chemical, in-vitro, and ex-vivo evaluations. The 23 FFD assessed the impact of total lipid amount, surfactant amount, and sonication time on critical quality attributes such as particle size and % entrapment efficiency. In-vitrorelease testing (IVRT) validation was performed using vertical diffusion cells. IVRT, a compendial technique by pharmacopoeias, was for performing semi-solid formulations analysis. The optimized EBR-NLC 1% w/w was characterized for assay, organic impurities, amplitude sweep, viscosity, IVRT, ex-vivo permeation testing, and skin retention. The validated IVRT technique was meeting the acceptance criteria of regulatory guidelines. The results showed that in-vitro release, ex-vivo permeation, and skin retention were significantly higher (P < 0.05) for the optimized EBR-NLC 1% w/w formulation compared to the innovator formulation (EBERNET® Cream 1% w/w). Applying QbD principles systematically facilitated the successful development and optimization of an EBR-NLC 1% w/w. The optimized EBR-NLC 1% w/w formulation proved to be a viable alternative, showing stability for at least six months under conditions of 40°C/75% RH and 30°C/75% RH.

Introduction Optimization of pharmaceutical formulations requires advanced tools to ensure quality, safety, and efficacy. quality-by-design (QbD), introduced by the FDA, emphasizes understanding and controlling processes early in development. Advanced optimization methods, such as desirability, have surpassed traditional single-objective techniques. Others, such as weighted goal programming (WGP) offers unique advantages by integrating decision-maker preferences, enabling balanced solutions for complex drug delivery systems. This study applies WGP to optimize timolol (TM)-loaded nanoliposomes aligning with QbD principles. Methods The optimization process followed six steps: identifying factors and responses, developing a Design of Experiments (DoE) plan, defining ideal and anti-ideal points, setting aspiration levels, assigning relative weights, and applying WGP compared to desirability function. Minimized and balanced deviations from aspiration levels served as criteria for selecting the most robust optimization results. Six responses were analyzed: vesicle size ( z 1 ) , polydispersity index ( z 2 ) , zeta potential ( z 3 ) , deformability index ( z 4 ) , phosphorus content ( z 5 ) , and drug entrapment efficiency ( z 6 ) . Results WGP produced a more balanced formulation that simultaneously optimized multiple responses. By incorporating the importance of each response, the WGP approach improved control over size, colloidal stability, and drug entrapment, based on its mathematical formulation. Comparative analysis with the desirability function confirmed that WGP effectively addressed potential tradeoffs without oversimplifying conflicting objectives. Conclusions This case-study demonstrates WGP potential as an advanced multi-objective optimization tool for pharmaceutical applications, improving upon traditional methods in complex formulations. Its ability to harmonize multiple critical attributes in line with QbD highlights its value in developing high-quality pharmaceutical products.

INTRODUCTION. Currently, manufacturers of adeno-associated virus (AAV)-based gene therapy products are facing a number of systemic problems stemming from the difficulties in assessing the quality of medicinal products due to insufficient scientific data, limited experience, and imperfect regulatory requirements. However, a risk-based approach to assessing critical quality attributes (CQAs) within the the framework of Quality by Design (QbD) can ensure improved efficiency in the development and production of advanced therapy medicinal products.AIM. This study aimed to identify QbD-based CQAs and associated specifications for the development of AAV-based gene therapy products for Duchenne muscular dystrophy.DISCUSSION. This study involved an analysis of QbD-based approaches to the development of AAV production technologies. The authors substantiated a list of the main AAV characteristics and collated available data on their impact on patients in terms of the efficacy and safety of gene therapy products and, in particular, the immune response to treatment. Following a risk assessment, the authors identified a list of CQAs for AAVs. When developing an AAV production process, the authors determined specifications for AAV CQAs, including viral and infectious titres, the presence of replication-competent AAVs, the percentage of empty capsids, and residual impurities (proteins, plasmid DNA, and residual host-cell DNA). A comprehensive risk assessment was conducted to determine the quality target product profile for an AAV-based gene therapy product for Duchenne muscular dystrophy. The authors listed the CQAs, developed the basic requirements for the applicable analytical procedures, and established the CQA specifications for the gene therapy product.CONCLUSIONS. The use of QbD principles and risk-based approaches is an important step in CQA identification during the development of gene therapy products. The QbD methodology facilitates drafting new regulatory standards for the evaluation of the safety and efficacy of gene therapy products and helps with the development and commercial-scale manufacturing of such products.

The use of dissolving microneedle arrays (dMNA) for intradermal and transdermal drug delivery has been a growing trend in the field for the past decades. However, a lack of specific regulatory standards still hinders their clinical development and translation to market. It is also well-known that dMNA composition significantly impacts their performance, with each new formulation potentially presenting a challenge for developers, manufacturers and regulatory agencies. A systematic approach such as quality-by-design (QbD), which embeds quality from the very beginning of the product development process, allows the design and optimisation of a drug-loaded dMNA formulation with promising features. In this work, we defined the Quality Target Product Profile (QTPP) for lidocaine-loaded dMNA and optimised their composition through a sequential design of experiments (DoE) approach. The first step (DoE_1) confirmed the influence of all formulation components (PVP, PVA and sucrose) in the properties of the arrays and pre-optimised their settings for DoE_2. The array characterisation focused on previously defined critical quality attributes (drug content, dissolution time, mechanical strength, skin insertion and physical attributes). At its maximum desirability (85.15%), the optimised design space obtained in DoE_2 is predicted to produce Li-dMNA with high mechanical strength (< 10% needle height reduction), skin insertion (> 90% needle height) and Li-HCl loading (~ 5mg), good physical attributes and dissolving in a maximum of 60min. The flexible design space obtained allows for the production of dMNA that consistently meet the QTPP, reducing batch failure and end-product testing, which are common in the more rigid GMP approach. Overall, applying QbD principles to formulation development shows promise to increase product quality and facilitate translation of dMNA into the clinic.

Advancements in understanding human developmental biology have greatly facilitated the differentiation of diverse functional cell types from human pluripotent stem cells (hPSCs). As a result, over a hundred clinical studies have been initiated, with several cell therapy products progressing to late-stage clinical development. However, ensuring the safety, efficacy, and quality of these cellular therapies remains a significant challenge. This chapter presents a Quality by Design (QbD)-based product development strategy, using a PSC-derived mesenchymal stromal cell-like (MSC-like) therapy as a case study. Given the complex biological nature of these "living" therapeutic agents and the limited prior knowledge available, the approach aligns with ICH guidelines to define the Quality Target Product Profile (QTPP) and applies risk assessment tools, such as risk ranking and filtering, to identify Critical Quality Attributes (CQAs). Additionally, Critical Process Parameters (CPPs) are optimized to enhance manufacturing robustness, addressing challenges related to viability, purity, and aseptic processing. The study also highlights the importance of analytical method validation, raw material control, and lifecycle management in meeting regulatory standards. By integrating QbD principles into PSC-derived therapy development, this work provides a structured framework for producing high-quality cell therapy products and establishes a valuable precedent for future cell-based therapies.

This research is intended to examine the adoption of the Quality by Design (QbD) approach within a process industry setting, thereby leading to sustainable performance. Specifically, the purpose of this study is to explore the systematic integration of QbD principles into the design and control of distillation column controllers to improve product quality, operating efficiency, and sustainability. The study analyzes process variables and develops a robust system by leveraging statistical methods, quality management tools, and chemical and control engineering expertise. It is found that effective deployment of QbD methodology in process engineering requires a robust and systematic approach. Additionally, we determined that Quality Target Product Profile (QTPP) and Critical Quality Attributes (CQA) are important components for effective deployment and sustainment of QbD to achieve Sustainable Development Goals for the industries. Moreover, it is observed that including noise and control parameters is essential during the project’s design phase. This study offers a systematic approach to implementing QM in industrial process design. The study highlights the potential of QbD in control engineering to enhance industrial processes and contribute to sustainability.

Optimizing extrusion processes and understanding conformational changes in itraconazole amorphous solid dispersions using in-line UV–Vis spectroscopy and QbD principles

ABSTRACT Clinical trials have shown that combining a modest dose of Rivaroxaban (RIV) with Aspirin (ASP) is more effective than ASP alone in reducing cardiovascular mortality, myocardial infarction, and stroke. A high‐performance liquid chromatography method developed using Quality by Design (QbD) effectively estimates ASP and RIV for routine pharmaceutical analysis. The Plackett‐Burman design assessed key factors including organic phase volume, mobile phase pH, flow rate, temperature, and injection volume using an Ishikawa fish‐bone diagram for risk assessment. These parameters were then optimized via Box‐Behnken design in 15 trials. The method meets the International Council For Harmonization standards with a mobile phase ratio of 45:55 (v/v) acetonitrile: buffer (pH 3, adjusted with orthophosphoric acid), a flow rate of 1 mL/min, a column temperature of 30°C, and an injection volume of 20 µL. A Phenomenex Luna C18 column (250 × 4.6 mm, 5 µm particle size) with detection at 251 nm ultra‐violet wavelength. The retention times of 4.54 min for ASP and 5.81 min for RIV were obtained. Validation confirmed linear ranges: 40–200 µg/mL for ASP and 2–10 µg/mL for RIV, with excellent recovery rates (ASP: 99.30%–101.9%; RIV: 98.52%–101.84%). Developed using QbD principles, the method proved specific, accurate, precise, sensitive, and robust for routine quality control of both compounds in pharmaceutical formulations.

Introduction: The aims of this study were to determine and govern critical sources of variability in the process and to investigate the effect of excipients and processing conditions on the critical quality attributes (CQAs) of isoniazid (INZ) crystal agglomerates using a quality-bydesign (QbD) paradigm along with risk-based approach. Methods: The QbD paradigm is thoroughly exercised to determine the CQAs and to, link these CQAs to agglomerate properties and to recognize potentially significant input variables. Considering this, a risk assessment tool was executed with different excipients and processing conditions to define their effect on CQAs of agglomerates. Potential risk factors were recognized using an Ishikawa diagram and then investigated using principal component analysis and an artificial neural network. After that, all agglomerates were optimized using the design of the experiment. Results: The optimized INZ crystal agglomerates were characterized using various tools such as X-ray diffractometry, Fourier transform infrared spectroscopy, scanning electron microscopy, differential scanning calorimetry, gas chromatography, and stability testing. These characterizations confirmed that INZ did not undergo any structural modifications in the agglomerates. Conclusion: The results indicate that the QbD principles and risk assessment tool provide a valuable means to fully understand directly compressible INZ crystal agglomerates, which could be considered an exceptional alternative to the granulation method.

The current trend in pharmaceutical process development and optimization is known as quality by design (QbD). This methodical approach is founded on the concept that quality should be integrated into the product from the start, rather than assessed after manufacture. In this study, we demonstrate methods to improve the process parameters for making tri-layer tablets by applying QbD principles in a methodical way. Dependent elements with high to medium impact on the process. According to the study’s results, all systems showed signs of instant release. It appears that the structure of each formulation and the properties of the polymer utilized have a significant impact on the rate and mechanism of drug release. Substantial medication release is observed in the three-layer formulations. Both the pace and mechanism of drug release are significantly affected by the tablet’s geometrical features and structure, as well as the weight and thickness of the barrier layers. While data suggested that Fickian diffusion was primarily responsible for drug release in two-layer tablets, three-layer tablets showed signs of either anomalous diffusion or erosion/relaxation. Therefore, the drug was stabilized as immediate-release directly compressible triple layered Average core weight of tri-layered tablets (mg) was 1000 for ibuprofen, 100 for placebo and 200 for ranitidine, thickness (mm) was 9.00 ± 0.3, hardness (N) was 110 ± 25, core tablet weight was 1300.000 ± 2% mg and coated tablet weight was 1325 mg. Using other moisture-sensitive medications. This study provides the foundation for future research into optimizing the properties of tablets made by direct compression.

This study emphasizes the crucial role of Quality by Design (QbD) in developing pharmaceutical procedures, particularly in risk assessment. It demonstrates how QbD principles were applied to create a precise and effective HPLC method for Silymarin Tablets, ensuring consistent quality within specified criteria. The optimized method, developed using a Design of Experiment approach, employs a C18 column (150 mm x 4.6 mm, 5μm) with isocratic elution using a 95:25 ratio of acetonitrile to orthophosphoric acid buffer (pH 3). Peaks were detected using a PDA detector calibrated at 287 nm, with a flow rate of 1.0 mL/min. The column oven temperature was maintained at 25°C, and a 10 μL injection volume was used. Thorough validation, adhering to USP &lt;1225&gt; and ICH Q2 (R1) standards, ensures the method's reliability. Key factors such as accuracy, precision, robustness, limit of detection (LOD), and limit of quantification (LOQ) were comprehensively assessed. The method exhibits exceptional sensitivity, selectivity, efficiency, precision, accuracy, and cost-effectiveness, making it ideal for pharmaceutical analysis of Silymarin tablets. It has been validated to effectively differentiate between marketed products, including those closely resembling the original. This method is intended for routine quality control analysis in the pharmaceutical industry, highlighting its suitability and reliability for ongoing use.

The study aimed to systematically enhance the fabrication process of flurbiprofen-loaded bilosomes (FSB) using Quality by Design (QbD) principles and Design of Experiments (DOE). The objective was to develop an optimized formulation with improved entrapment efficiency and targeted drug delivery capabilities. The optimization process involved applying QbD principles and DOE to achieve the desired formulation characteristics. Superparamagnetic iron oxide nanoparticles (SPIONs) were incorporated to impart magnetic responsiveness. The size, entrapment efficiency, morphology, and in vitro release patterns of the FSB formulation were evaluated. Additionally, an in situ forming hydrogel incorporating FSB was developed, with its gelation time and drug release kinetics assessed. In vivo studies were conducted on osteoarthritic rats to evaluate the efficacy of the FSB-loaded hydrogel. The optimized FSB formulation yielded particles with a size of 453.60nm and an entrapment efficiency of 91.57%. The incorporation of SPIONs enhanced magnetic responsiveness. Morphological evaluations and in vitro release studies confirmed the structural integrity and sustained release characteristics of the FSB formulation. The in situ forming hydrogel exhibited a rapid gelation time of approximately 40 ± 1.8s and controlled drug release kinetics. In vivo studies demonstrated a 27.83% reduction in joint inflammation and an 85% improvement in locomotor activity in osteoarthritic rats treated with FSB-loaded hydrogel. This comprehensive investigation highlights the potential of FSB as a promising targeted drug delivery system for the effective management of osteoarthritis. The use of QbD and DOE in the formulation process, along with the integration of SPIONs, resulted in an optimized FSB formulation with enhanced entrapment efficiency and targeted delivery capabilities. The in situ forming hydrogel further supported the formulation's applicability for injectable applications, providing rapid gelation and sustained drug release. The in vivo results corroborate the formulation's efficacy, underscoring its potential for improving the treatment of osteoarthritis.

Background: Fenofibrate (FEN) is the FDA-approved drug used in the treatment of hyperlipidemia. FEN possesses limited bioavailability orally due to its low solubility. As a result, more frequent and larger doses are needed, which increases the likelihood of adverse effects. Objectives: This study aimed to develop and optimize polymeric nanoparticles loaded with Fenofibrate (FEN) using the solvent evaporation method. Method: A Quality by Design (QbD) approach was used to ensure the quality of the finished product by evaluating the impact of critical material attributes (CMAs) and critical process parameters (CPPs) on the critical quality attributes (CQAs) of nanoparticles. The impact of CMAs (quantity of polycaprolactone, % polyvinyl alcohol, and % sodium lauryl sulphate) on particle size and Drug Entrapment Efficiency (DEE) was studied using Box-Behnken Design. Results: The optimized nanoparticles have 246.5 ± 4.38 nm particle size and 77.53 ± 0.9% DEE. SEM and TEM were used to analyze the surface morphology of nanoparticles. Furthermore, In-Vitro drug release study of optimized formulation was performed to confirm the efficacy of the polymeric nanoparticles. Conclusion: The solvent evaporation method was utilized to effectively formulate FEN-loaded polymeric nanoparticles and optimized through QbD principles to achieve minimum particle size and maximum % DEE

Hydrophobic ion pairing (HIP) complexation was found to be an efficient approach in modulating the release and enhancing the stability and encapsulation of hydrophilic macromolecules such as proteins in hydrophobic nano/microcarriers. The present work strives to develop and optimize the preparation of the HIP complex of the antimicrobial enzyme lysozyme (LYZ) with the ion-pairing agent (IPA) sodium dodecyl sulphate (SDS) relying on the quality-by-design (QbD) approach. The quality target product profile (QTPP) includes the achievement of maximal lipophilicity in a reversible manner to enable the maintenance of biological activity. The related critical quality attributes (CQAs) were defined as complexation efficacy, complex stability, enzyme recovery and activity. Three risk assessment (RA) tools were used to identify and rank the critical process parameters (CPPs) and critical material attributes (CMAs). From this assessment, the pH of the medium, LYZ:SDS molar ratio and drying conditions were determined as high-risk factors that need to be investigated. To the best of our knowledge, for the first time, electrostatic titration was used as a smart approach to determine the optimum molar ratio at different pH values. Based on the predefined CQAs, pH 8 with an LYZ/SDS molar ratio of 1:8 was found to be the optimal condition for complexation efficiency and recovery (%) of a biologically active enzyme. A cost-effective drying process based on a ventilated oven was developed, which resulted in complex qualities comparable to those obtained by the commonly used freeze-drying method. In a nutshell, the optimum conditions for the preparation of the LYZ/SDS HIP complex were efficiently facilitated by the rational application of QbD principles and the utilization of efficient electrostatic titration and ventilated oven-drying methods.