Int J Chem Res, Vol 10, Issue 2, 6-11Review Article


A REVIEW ON THE ROLE OF KINETICS IN ELUCIDATING THE MECHANISM OF BIOLOGICALLY ACTIVE MOLECULES

FAKEHA MOHAMMED REHAN SHAIKH, ASHISH SAMBHAJI UZGARE*

Department of Chemistry, Faculty of Science, Wilson College (Autonomous), Mumbai-400007, Maharashtra, India
*Corresponding author: Ashish Sambhaji Uzgare; *Email: [email protected]

Received: 25 Aug 2025 Revised and Accepted: 12 Dec 2025

ABSTRACT

Kinetic analysis plays a crucial role in elucidating the behavior of biologically active molecules, providing detailed insights into their interaction mechanisms, stability, and functional dynamics. This review underscores the importance of kinetic studies in enzyme catalysis, drug-target binding, and the behavior of lectins and glycoproteins. By examining kinetic parameters such as rate constants for association and dissociation, researchers can gain valuable information about the potency of inhibitors, the thermal stability of lectins, and the binding kinetics of glycoproteins. These parameters reveal essential aspects of molecular interactions, helping to assess the efficiency and specificity of therapeutic agents. Moreover, integrating kinetic data with genomic and proteomic information has propelled the field of systems biology, offering a more holistic understanding of molecular processes. Kinetic studies are also important for optimizing therapeutic strategies, as they provide insights into drug efficacy and resistance mechanisms. Additionally, they enhance the design of bioanalytical tools, leading to more accurate and efficient techniques for studying complex biological systems.

Keywords: Kinetics, Biologically active molecule, Reactions

© 2026 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)
DOI: http://dx.doi.org/10.22159/ijcr.2026v10i2.308 Journal homepage: https://ijcr.info/index.php/journal

INTRODUCTION

Kinetic studies focus on chemical steps designed to determine an appropriate mechanism model, correlating it with experimental data to find the best fit that closely represents the reaction rate and characterizes the chemical process [1]. Kinetic studies are essential for gaining a deeper understanding of the leaching process's nature and mechanism. Their purpose is to identify the most suitable kinetic model, extract relevant kinetic parameters, and provide insights that can aid in plant design, optimization of operating conditions, real-time control, and maximizing metal recovery [2]. The field of reaction kinetics focuses on understanding the rate dynamics in chemical and biological systems. By experimentally determining reaction rates, it is possible to estimate rate constants and enhance our comprehension of biological systems modeled by reaction-diffusion processes [3]. The exploration of kinetic principles in the study of biologically active molecules has been crucial for drug development, providing insights into the effectiveness, specificity, and binding affinity of compounds. Enzyme kinetics, in particular, has significantly enhanced our understanding of how enzymes catalyze biochemical reactions, revealing key aspects of enzyme-substrate interactions. This knowledge has been essential for designing inhibitors with therapeutic potential [4]. Kinetic studies of enzymes constitute a significant area of research and have greatly enhanced our understanding of enzyme mechanisms [5]. Moreover, kinetics can help predict the behavior of biologically active molecules under various physiological conditions, offering insights into their potential for therapeutic use. Techniques such as surface plasmon resonance (SPR), isothermal titration calorimetry (ITC), and fluorescence anisotropy have become widely used to assess the kinetic parameters of drug-target interactions [6]. The combination of kinetic models with genomic and proteomic data has transformed systems biology, providing a more comprehensive understanding of biological processes at the molecular scale [7].

A review of the scientific literature on kinetics aimed to focus on research related to kinetic studies in elucidating the mechanisms of biologically active molecules. Data were gathered from various online scientific literature sources, including databases such as Google Scholar, Web of Science, Scopus, and Science Direct. Specific keywords related to kinetics studies on different biologically active molecules utilized in the search strategy. The study’s inclusion criteria involved selecting research that contained one or more of these keywords in the article titles, focusing on articles published in English-language journals. Total 50 articles were selected for review from 1986 to 2025.

Role of kinetics

Table 1 indicates role of kinetics in biologically active molecule.

Table 1: Role of kinetics

Author(s) Year Study focus References
Bachurin et al. 1993 Kinetic characteristics of reversible enzyme inhibition (AChE) [8]
Weggen et al. 2023 Scale-up of ADC manufacturing using kinetics and CFD simulations [9]
Yukawa et al. 1967 Kinetic study of Wolff rearrangement of α-diazoacetophenone [10]
Hoebeke et al. 1978 Kinetics of Lens culinaris lectin binding to rabbit erythrocytes [12]
Shnyrov et al. 1996 First-order kinetics on lentil lectin, temperature-dependent rate constants, activation energy calculated using the Arrhenius equation [14]
Kitano et al. 2009 Binding kinetics of ConA to mannose residues in glycopolymer brushes [15]
Ralin, D. W. et al. 2008 kinetics of glycoprotein–lectin interactions were studied using label-free internal reflection ellipsometry (LFIRE™) [16]
Kato et al. 1997 Kinetic analysis of lectin-reactive α-fetoprotein (AFP) with different lectins [18]
Hooper et al. 2022 Kinetics of multivalent lectin-glycan interactions using glycan-QDs [19]
Nantwi et al. 1997 Kinetics of lectin binding to human buccal cells [20]
Shinohara et al. 1997 Kinetics of lectin-oligosaccharide interactions [21]
Lebed et al. 2007 Kinetics of ConA-Carboxypeptidase Y binding [22]
Tholen et al. 2025 Lectin-PAINT for multiplexed visualization of the glycocalyx [23]
Walter et al. 1999 Kinetics of lectin levels in Penaeus vanammei after antigen exposure [24]
Goumenou et al. 2021 Glycosylation's impact on protein activity and stability [25]
Swamy et al. 1986 Kinetics of N-dansylgalactosamine binding to SBA using stopped-flow spectrofluorimetry [26]
Roy et al., 2011 Kinetics of lectin binding influenced by hydrodynamic shear forces [27]
Fernandez-Poza et al. 2021 Kinetics of recombinant prokaryotic lectins (RPLs) binding to glycan residues [28]
Damian et al. 2005 Binding kinetics of Xerocomus chrysenteron lectin (XCL) [31]
Sakonwan et al. 2020 LecA/PA-IL galactose-binding lectin kinetics [33]
Khan et al. 2013 Kinetic studies, temperature and pH analysis of Bananaa lectin [34]
Bhangare et al. 2022 Degradation kinetics of pharmaceuticals [35]
Abebrese et al. 2011 Kinetics of oxygen atom transfer from trans-dioxoruthenium(VI) [36]
Wakelin et al. 1987 Dissociation kinetics of DNA complexes with antitumor drugs [37]
AlRashidi et al. 2024 Kinetic studies, molecular docking simulations [38]
Weerawardhana et al. 2023 Kinetic studies, activation parameters [39]
Habibi-Khorassani et al. 2008 Kinetics of reaction between triphenylphosphine and acetylenedicarboxylates [40]
Grunwald and Krueger 2008 Bioinformatics in enzyme kinetics [41]
Sadutto et al. 2023 HPLC, pseudo-first-order rate constant measurements [42]
Mustafa et al. 2023 Kinetic analysis, molecular dynamics and docking simulations [43]
Banerjee et al. 2014 Kinetic properties of adenylosuccinate lyase (ASL) from Mycobacteria [44]
Sigmundsson et al. 2002 Kinetics of biomolecular interactions using SPR [45]
Basha et al. 2013 Kinetics of oxyhemoglobin oxidation by thiosemicarbazone complexes [46]
Tarhouchi et al. 2022 Kinetic and thermodynamic analysis, diffusion studies [47]
Sung and Nesbitt 2020 DNA hybridization kinetics under extreme pressures [48]
Van Oijen 2011 Single-molecule fluorescence, real-time binding event tracking [49]

The study by Bachurin et al. assesses the kinetic characteristics of various mechanisms of reversible enzyme inhibition, focusing on their efficacy in relation to substrate concentration. The research examines the inhibition of acetylcholinesterase (AChE) by phenylpyridinium derivatives and their effects on animals. Key findings include the significant impact of inhibition mechanisms on enzyme efficacy and the differential toxic effects of MPP, EPP, and BPP. The study highlights the importance of kinetic parameters in understanding and optimizing the physiological activity of these compounds [8].

Weggen, et al. A study investigates the scale-up of antibody-drug conjugate (ADC) manufacturing using experimental kinetics and CFD simulations. It shows that internal stirrer mixing provides more robust kinetics than external mixing devices. Simulations of large-scale reactors reveal that mixing is sufficient at all scales, with deviations occurring mainly during payload addition. The findings highlight the use of kinetic models to optimize ADC production without large-scale experiments [9].

Kinetic studies of the Wolff rearrangement of α-diazoacetophenone were conducted by Yukawa Y et al. in t-butyl alcohol, measuring nitrogen evolution using silver benzoate in triethylamine as a catalyst. The reaction followed pseudo-first-order kinetics with an induction period. The effects of silver benzoate and triethylamine concentrations on the reaction rate were examined, with silver ions being consumed only in the reaction's initial stage. Based on these findings, a new mechanism was proposed, suggesting that the active catalyst entity is a silver ion-triethylamine complex [10]. I He et al. study the thermal stability of lectin from black turtle bean was analyzed using first-order reaction kinetics. Kinetic data showed that the inactivation of lectin followed first-order kinetics, with an activation energy of 78.80 kJ/mol. This indicates the lectin's thermal sensitivity and hydrophobic stability. The results underscore the importance of temperature and treatment time in determining the thermal stability of the lectin [11].

Hoebeke, et al., in conducted Kinetic studies on Lens culinaris lectin binding to rabbit erythrocytes, yielding association and dissociation rate constants of 3–10 x 10³ M⁻¹ s⁻¹ and 3–33 x 10⁻⁴ M⁻¹ s⁻¹, respectively, over a temperature range of 5 to 37 °C. The activation energies were 7 kcal/mol for the forward reaction and 13 kcal/mol for the reverse. These kinetic parameters were consistent with equilibrium findings, showing the binding process to be exothermic (ΔH =-4.6 kcal/mol) and associated with positive entropy (ΔS =+15 eu) [12].

Dam et al. examined the binding and cross-linking of concanavalin A (ConA) and Dioclea grandiflora lectin (DGL) with synthetic divalent carbohydrates. Results showed that carbohydrates with flexible spacers between terminal Man residues had higher affinities for both lectins. Kinetic and electron microscopy studies revealed differences in cross-linking behavior, with increased spacer length leading to the loss of organized cross-linked lattices. These findings highlight the impact of spacer flexibility on lectin-carbohydrate interactions [13].

Shnyrov et al. studied the irreversible thermal denaturation of lentil lectin at pH 7.4 was analyzed using a first-order kinetic model. The temperature-dependent rate constant, described by the Arrhenius equation, was calculated along with the activation energy. Results from differential scanning calorimetry, fluorescence, and hemagglutination assays supported the two-state kinetic mechanism [14].

Kitano H. et al., focus on the binding kinetics of Concanavalin A (Con A) to mannose residues in a disulfide-carrying glycopolymer brush (DT-PMEMan) were studied using UV–vis spectroscopy and localized surface plasmon resonance. Con a exhibited concentration-dependent binding, with a significantly higher apparent association constant compared to small sugars, attributed to the cluster effect. The glycopolymer brush demonstrated a detection limit below 5 nM, highlighting the enhanced binding kinetics and sensitivity of the system [15].

Ralin et al. study, kinetics of glycoprotein–lectin interactions were studied using label-free internal reflection ellipsometry (LFIRE™), showing improved binding with multi-lectin strategies. LFIRE™ offers a cost-effective, high-throughput solution for real-time kinetic screening, enabling the rapid identification of glycoprotein biomarkers and providing insights into glycan functionality in cellular health and disease [16].

Sanadi, A. R., and Surolia, A. Study Coccinia indica agglutinin (CIA) is a chitooligosaccharide-specific lectin with two binding sites. Tryptophan involvement in binding was confirmed through quenching and chemical modification studies. Thermodynamic analysis revealed that binding is entropically driven, with the nonreducing sugar protruding from the binding pocket. Stopped-flow kinetic studies indicated that the binding process follows a one-step mechanism [17].

The study by Kato et al. focused on the affinity and kinetic analyses of lectin-reactive α-fetoprotein (AFP) using a surface plasmon resonance (SPR) biosensor. The researchers immobilized human AFP from hepatocellular carcinoma (HCC) patients onto a sensor chip and monitored its interaction with three lectins: Lens culinaris agglutinin (LCA), concanavalin A (Con-A), and erythroagglutinating phytohemagglutinin (E-PHA). Real-time SPR monitoring revealed distinct variations in the association (k_ass) and dissociation (k_diss) rate constants for each lectin-AFP interaction. This SPR-based method offers a rapid and efficient technique for evaluating lectin-AFP interactions and provides detailed kinetic insights into their binding behavior [18].

Hooper et al. introduces glycan-functionalized quantum dots (glycan-QDs) to probe multivalent lectin-glycan interactions (MLGIs) and their kinetics. Using the lectins DC-SIGN and DC-SIGNR, the researchers observed that DC-SIGN binds a single glycan-QD, while DC-SIGNR cross-links multiple QDs. Kinetic analysis revealed that DC-SIGN binding follows a single, fast second-order association rate, while DC-SIGNR exhibits two distinct association phases: a rapid initial binding followed by a slower secondary interaction. This highlights the potential of glycan-QDs for studying the kinetics of complex, multivalent interactions [19].

Nantwi et al. investigated the kinetics of lectin binding to human buccal cells, showing significant binding within 20 seconds, particularly for lectins from Pisum sativum and Arachis hypogaea. The study found that lectin binding was reduced when buccal cells were pre-treated with secretor or non-secretor saliva, although once bound, lectins were not displaced by saliva. These findings highlight the rapid kinetics of lectin binding to buccal cells and the influence of saliva on binding strength [20].

Shinohara et al. examined the kinetics of lectin-oligosaccharide interactions using a solid-phase assay. They found that increasing oligosaccharide density decreased both the association (k_ass) and dissociation (k_diss) rate constants. Lectin oligomerization enhanced binding by reducing k_diss. These results highlight the role of mass transport and rebinding in modulating lectin kinetics [21].

Lebed et al. used atomic force microscopy (AFM) and quartz crystal microbalance (QCM) to study the kinetics of concanavalin A-carboxypeptidase Y binding. Both methods yielded similar dissociation rate constants (k_diss), with QCM also providing the association rate (k_ass = 5.6±0.1 × 10⁴ M⁻¹ s⁻¹) and equilibrium constant (K_a = 0.59±0.01 × 10⁶ M⁻¹). AFM revealed the energy barrier for dissociation (r = 2.29±0.04 Å). The study highlights the complementary use of AFM and QCM for kinetic analysis of molecular interactions [22].

Tholen et al. introduced Lectin-PAINT, a super-resolution imaging technique for multiplexed, live-cell visualization of the glycocalyx at the single-cell and single-molecule levels. By leveraging lectin binding to specific carbohydrates, Lectin-PAINT enables nanoscale mapping of glycans with high resolution. Using an extensive lectin library, the method allows for the extraction of over 350 glycosylation parameters per cell, creating a "glycotype." This approach was applied to classify cancer cell types, revealing significant glycocalyx heterogeneity and variability, with potential for advancing understanding of glycocalyx changes in disease [23].

Walter et al. studied lectin level kinetics in Penaeus vanammei after antigen exposure. They observed that lectin levels recovered more quickly after a second challenge, indicating an adaptive immune response. This pattern could serve as a potential marker for evaluating disease resistance in shrimp strains for aquaculture [24].

Goumenou, A., Delaunay, N., and Pichon, V. Glycosylation significantly impacts protein activity and stability. Despite advances, fully characterizing glycosylation is challenging due to complex mixtures in biological samples. Lectin-based affinity sorbents are commonly used for selective enrichment of glycosylated proteins or peptides, aiding in their analysis. The review highlights recent progress in lectin affinity techniques, enabling faster and more automated glycosylation analysis, with a focus on kinetic aspects of protein-enrichment interactions [25].

Swamy et al. studied the kinetics of N-dansylgalactosamine binding to soy-bean agglutinin (SBA) using stopped-flow spectrofluorimetry. The binding followed a single-step mechanism with association and dissociation rate constants of k+1 = 2.4 × 10⁵ M⁻¹·s⁻¹ and k−1 = 0.2 s⁻¹ at 20 °C. The kinetic data indicated an enthalpy-driven process [26].

Roy et al. demonstrate that the reaction kinetics of lectins, such as Concanavalin A and Abrus Agglutinin, are influenced by hydrodynamic shear forces. Binding efficiency declines beyond a critical shear rate, indicating that association and dissociation constants depend on hydrodynamic conditions. This suggests that strong shear forces can distort lectin structure, altering their kinetics. The study emphasizes the importance of considering hydrodynamics in microfluidics-based assays [27].

In their 2021 study, Fernandez-Poza et al. examine the development of novel recombinant prokaryotic lectins (RPLs) through directed evolution, focusing on their binding kinetics to specific glycan residues (α-mannose, β-galactose, fucose, and sialic acid). Using bio-layer interferometry (BLI), the study compares the binding activity and specificity of RPLs to plant-derived lectins. RPLs showed superior binding in most cases, with significantly better limits of detection (LoDs) for glycoprotein interactions [28].

Murthy and Jayaraman performed a kinetic analysis of the T-antigen's interaction with peanut agglutinin lectin, employing surface plasmon resonance. Their findings showed that the initial antigen-lectin binding step had a fast association rate and a slow dissociation rate. In contrast, subsequent binding events exhibited slower kinetics. The study also compared these kinetics with other glycoside derivatives, highlighting differences in the interaction rates [29].

Vico et al. investigated the kinetics of vesicle aggregation in a system with β-cyclodextrin vesicles decorated with maltose-and lactose-adamantane conjugates interacting with lectins. They found that aggregation rates were proportional to lectin and vesicle concentrations, with each lectin requiring a specific carbohydrate density for optimal binding. This study provides insights into multivalent interactions and ligand-receptor kinetics at interfaces [30].

Damian et al. studied the binding kinetics of Xerocomus chrysenteron lectin (XCL) using isothermal titration calorimetry. They found that XCL binds N-acetylgalactosamine with a dissociation constant (Kd) of 5.2 μM at 27 °C, and the binding affinity increases at lower temperatures. The dissociation constant for the Thomsen-Friedenreich antigen was 500 times higher (Kd = 0.94 μM), suggesting XCL's important role in recognizing specific glycan motifs [31].

Zaree et al. investigated the binding kinetics of mono-and divalent ligands to Pseudomonas aeruginosa LecA using multiple techniques. They found that the divalent ligand had a low-nanomolar affinity and a residence time of about 7 h, with no strong binding to related lectin tetramers. These findings provide key insights into the binding kinetics and chelation-based interactions of the ligand with LecA [32].

Sakonwan et al. study LecA/PA-IL is a galactose-binding lectin from Pseudomonas aeruginosa that binds α-galactose on glycosphingolipids, playing a role in cell adhesion and bacterial internalization. It also exhibits direct toxic activity. Kinetic studies, including surface plasmon resonance assays and structural analyses, can be used to identify high-affinity inhibitors for this lectin [33].

Khan et al. studied the pH-induced monomerization of banana lectin (BL), showing that at pH 2.0, BL transitions to a molten globule-like state. The monomeric form exhibited greater resistance to chemical denaturation than the native dimer, with higher Cm and ΔG values. Additionally, the monomer retained secondary structure up to 95 °C, while the dimer's Tm was 77 °C, indicating superior stability of the monomer under denaturing conditions [34].

Bhangare et al. highlight the importance of degradation kinetic studies in determining the shelf life of pharmaceuticals under various conditions. The article discusses the lack of systematic strategies and regulatory guidelines for such studies and emphasizes the need for proper selection of variables to ensure accurate kinetic data. It provides a comprehensive approach to degradation kinetics, including advanced models, offering valuable insights for researchers in stability analysis to establish optimal storage conditions for drugs [35].

Abebrese et al. studied the kinetics of oxygen atom transfer from three trans-dioxoruthenium(VI) porphyrins to organic sulfides, focusing on sulfoxidation reactions with thioanisoles. The second-order rate constants for these reactions ranged from 8 to 60 M⁻¹ s⁻¹, significantly faster than those for alkene epoxidations or C–H bond oxidations. Reactivity followed the order 2a<2b<2c, consistent with the electron-withdrawing and steric effects of the porphyrin groups. Kinetic analysis suggested a concerted oxygen transfer mechanism, and competition reactions indicated a multiple oxidant model for sulfoxidation [36].

Wakelin et al. studied the dissociation kinetics of DNA complexes with the antitumor drug N-[2-(dimethylamino)ethyl]-9-aminoacridine-4-carboxamide and its derivatives. They found a fourth, stable binding mode in derivatives with a 4-CONH(CH2)2NR1R2 side chain, which correlated with better antitumor activity and selective binding to GC-rich DNA. The results support a molecular model where the drug intercalates into the narrow groove of DNA, with key hydrogen bonds contributing to its biological activity [37].

Al Rashidi et al. synthesized imidazo-isoxazole derivatives and evaluated their α-amylase and α-glucosidase inhibitory activity. Compound 5f exhibited the best inhibitory potency (IC50 of 26.67±1.25 μM and 39.12±1.83 μM, respectively) and was identified as a competitive inhibitor for both enzymes, similar to acarbose. Kinetic studies indicated that substitutions on the R1 and R2 groups of the thiourea/urea moiety influenced activity, with 5f showing significant binding affinity in molecular docking and stability in molecular dynamics simulations, supporting its potential as a lead compound for diabetes management [38].

Weerawardhana et al. synthesized [2Fe–2E] complexes (E = S, Se) with an N-alkyl, N′-aryl-β-diketiminate ligand and studied their electron transfer properties. The [2Fe–2S] complex exhibited an unusually long Fe⋯Fe distance, while the [2Fe–2Se] complex had a more typical Fe⋯Fe distance. Kinetic studies and activation parameters provided insights into the formation mechanism of the [2Fe–2S] compound. The study also explored the atom transfer reactivity of the [2Fe–2S] cluster to investigate its sulfur-transfer mechanism [39].

Habibi-Khorassani et al. investigated the kinetics of the reaction between triphenylphosphine and dialkyl acetylenedicarboxylates in the presence of NH-acid (benzoxazoline), using UV spectrophotometry to monitor the reaction. The reaction followed second-order kinetics, and the second-order rate constants (k₂) were calculated, with Arrhenius analysis yielding activation energy values. The study also examined the effects of solvent, reactant structure, and reactant concentration on the reaction rate. The proposed mechanism was confirmed, with the first step (k₂) identified as the rate-determining step. Additionally, isomer stability (Z or E) was analyzed using AIM theory [40].

Grunwald and Krueger developed a bioinformatics laboratory exercise that complements traditional alkaline phosphatase kinetics experiments in biochemistry courses. In this exercise, students use the Protein Explorer program to analyze the enzyme’s structure, specifically its active-site residues, and propose their functions. By linking kinetic data to the enzyme’s mechanism, students identify the roles of active-site residues in catalysis. A paired t-test of pre-and post-exercise assessments demonstrated that the exercise significantly improved students' ability to use kinetic data to determine the catalytic function of amino acids in the active site [41].

Sadutto et al. provided valuable kinetic data for understanding the factors influencing the stability and conversion of imine and enamine tautomers. These findings could help optimize conditions for maintaining the stereochemical integrity of similar bioactive compounds during synthesis [42].

Mustafa et al. conducted kinetic studies to evaluate the inhibitory effect of novel quinolinyl-iminothiazolines on alkaline phosphatase. The kinetic analysis revealed that the compound N-benzamide quinolinyl iminothiazoline (6g) exhibited the most potent inhibition, with an IC50 value of 0.337 ± 0.015 µM, significantly outperforming the standard KH2PO4 (IC50 = 5.245 ± 0.477 µM). The kinetic data, supported by molecular dynamics and docking studies, confirmed the strong inhibitory potential of 6g against alkaline phosphatase, indicating its promising role as a lead molecule for further drug development [43].

Banerjee et al. in their study on adenylosuccinate lyase (ASL) from Mycobacterium smegmatis (MsASL) and Mycobacterium tuberculosis (MtbASL), the enzyme's kinetic properties were evaluated. X-ray crystallography revealed that MsASL has a partially ordered C3 loop, with His149 and either Lys285 or Ser279 likely functioning as the catalytic acid and base. The enzymes from mycobacteria showed lower catalytic activity compared to other organisms' ASLs, which could be attributed to the Arg319Gly substitution and reduced flexibility in the C3 loop. These findings align with the slow growth rate of Mycobacteria and their reliance on alternative purine salvage pathways [44].

Sigmundsson et al. developed a method for accurately determining kinetic rate constants of biomolecular interactions using surface plasmon resonance (SPR) technology. The approach addresses mass transport limitations and does not require standards of known concentrations, relying instead on molecular mass. By analyzing binding curves obtained at varying flow rates, they introduced an analytical solution to the differential equations for 1:1 bimolecular complex formation. This method allows the precise determination of association and dissociation rate constants, as demonstrated by experiments with tyrosine-phosphorylated recombinant proteins and anti-phosphotyrosine antibodies, achieving picomolar sensitivity. The results showed strong agreement with traditional methods, highlighting the effectiveness of SPR for kinetic studies [45].

Basha et al. studied the kinetics of oxyhemoglobin oxidation by iron(III) thiosemicarbazone complexes, observing a two-step process involving the sequential oxidation of oxyheme cofactors in the α and β chains. Kinetic differences across complexes were attributed to steric and hydrogen-bonding effects in the thiosemicarbazone ligand series (HDpxxT), influencing the reaction rate [46].

Tarhouchi et al. investigated the kinetics of paracetamol diffusion across polymer membranes, specifically polymer inclusion membranes (PIM) and grafted polymer membranes (GPM) containing gluconic acid. The study showed that the PVA–GA membrane outperformed GPM–GA, with diffusion coefficients of 41.807 and 31.211 cm²/s, respectively. Kinetic and thermodynamic analysis indicated that the process was controlled by kinetic factors rather than energy, as evidenced by low activation energy and enthalpy values. The extraction process was most efficient at pH 1, providing a sustainable method for recovering paracetamol from aqueous solutions [47].

Sung and Nesbit studied DNA hybridization kinetics under extreme pressures, finding that pressure slowed folding and accelerated unfolding. Van’t Hoff analysis showed an increase in the DNA-solvent volume during folding. Sodium ions ([Na+]) stabilized DNA and reduced pressure dependence on folding kinetics, with higher [Na+] decreasing activation volumes for dehybridization, suggesting a shift to a more folded transition state [48].

Van Oijen discusses the use of single-molecule fluorescence techniques to explore the kinetics of biomolecular interactions. These methods enable real-time tracking of binding events by labeling molecules with fluorescent tags. While significant advancements have been made, the requirement for low, nanomolar concentrations of molecules remains a challenge for studying intermolecular interactions [49].

CONCLUSION

Kinetic studies are essential for advancing our understanding of biologically active molecules and their interactions. By examining kinetic parameters, such as rate constants for association and dissociation, researchers gain valuable insights into the potency and specificity of inhibitors. Kinetic analysis is an indispensable tool in both basic and applied biomedical research, providing a detailed understanding of molecular interactions. It plays a key role in optimizing therapeutic strategies and refining bioanalytical techniques. As the field evolves, the integration of kinetic studies with advanced technologies and interdisciplinary approaches will drive further breakthroughs in our understanding of biological systems. This, in turn, will facilitate the development of novel therapeutics and enhance the precision of biomedical interventions.

ACKNOWLEDGEMENT

The authors would like to acknowledge the valuable contributions of fellow researchers, whose work is cited in the references section, for their insightful publications that have informed and enriched this review paper. The table made by the authors themselves doesn’t copy from any sources.

FUNDING

Nil

AUTHORS CONTRIBUTIONS

Manuscript written by Fakeha Shaikh under the guidance of Ashish Uzgare. Fig. and table made by author themselves doesn’t copy from any sources.

CONFLICT OF INTERESTS

Authors declare no conflict of interest

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