Introduction: The purpose of this study was to evaluate the feasibility of using gel electrophoresis combined with autoradiographic image analysis for quantitating protein drug concentrations in biological fluid for pharmacokinetic studies.
Methods: Protein drugs were iodinated using the Iodogen reagent and injected into Sprague-Dawley rats for pharmacokinetic evaluation. Serum samples were analyzed using trichloroacetic acid (TCA)-precipitable counts or sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Commercially available precasted Bis-Tris gradient gels were used for SDS-PAGE. Autoradiography of gel samples was performed using phosphoimager and quantitated using the ImageQuant software.
Results: The maximum loading volume for protein drugs with molecular weight close to that of albumin ( approximately 70 kDa) was about 1 microl, whereas for protein drugs with larger or smaller molecular weight (i.e., >80 or <40 kDa), the maximum loading volume was up to 20 microl/lane. The optimal exposure time was about 18 h or overnight. Standard curves were constructed using serially diluted dosing solution, which was linear over a 10-fold concentration range with a correlation coefficient of.98. Comparing to the soluble human interleukin-13 receptor (shIL-13R) pharmacokinetic profiles from TCA-precipitable counts, the quantitative gel analysis revealed lower concentrations at later time points and a lower bioavailability from intraperitoneal injection.
Discussion: This study provided the first systemic evaluation of gel electrophoresis technology for quantitative protein drug determination in serum and its application in pharmacokinetic studies. The combination of gel electrophoresis with autoradiographic image analysis provided accurate and specific quantitation results. The overnight turnover time allowed routine application in preclinical pharmacokinetic studies.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/s1056-8719(02)00203-4 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Pan-cancer analysis of Arp2/3 complex subunits: focusing on ARPC1A's role and validating the ARPC1A/c-Myc axis in non-small cell lung cancer.
Front Immunol
January 2025
Department of Laboratory Medicine, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China.
Background: The Arp2/3 complex is a key regulator of tumor metastasis, and targeting its subunits offers potential for anti-metastatic therapy. However, the expression profiles, prognostic relevance, and diagnostic value of its subunits across cancers remain poorly understood. This study aims to investigate the clinical relevance of Arp2/3 complex subunits, particularly ARPC1A, in pan-cancer, and to further analyze the potential biological mechanisms of ARPC1A, as well as its association with immune infiltration and chemotherapy drug sensitivity.
View Article and Find Full Text PDFType IV collagen derived non-collagenous domain α6 (IV) NC1 and its derivative fragments inhibit endothelial cell proliferation and attenuates chorioallantoic membrane angiogenesis.
Cytotechnology
April 2025
Department of Genetics, Osmania University, Hyderabad, Telangana State India.
Targeting tumor angiogenesis with safe endogenous protein inhibitors is a promising therapeutic approach despite the plethora of the first line of emerging chemotherapeutic drugs. The extracellular matrix network in the blood vessel basement membrane and growth factors released from endothelial and tumor cells promote the neovascularization which supports the tumor growth. Contrastingly, small cleaved cryptic fragments of the C-terminal non collagenous domains of the same basement membrane display antiangiogenic effect.
View Article and Find Full Text PDFComputational identification of novel natural inhibitors against triple mutant DNA gyrase A in fluoroquinolone-resistant Typhimurium.
Biochem Biophys Rep
March 2025
Department of Integrative Biology, School of Biosciences and Technology, Vellore Institute of Technology (VIT), Vellore, 632014, Tamil Nadu, India.
The rising resistance to fluoroquinolones in Typhimurium poses a significant global health challenge. This computational research addresses the pressing need for new therapeutic drugs by utilizing various computational tools to identify potential natural compounds that can inhibit the triple mutant DNA gyrase subunit A enzyme, which is crucial in fluoroquinolone resistance. Initially, the three-dimensional structure of the wild-type DNA gyrase A protein was modeled using homology modeling, and followed by mutagenesis to create the clinically relevant triple mutant (SER83PHE, ASP87GLY, ALA119SER) DNA gyrase A protein structure.
View Article and Find Full Text PDFAmphotericin B tissue penetration and pharmacokinetics in healthy and -infected rats: insights from microdialysis and population modeling.
Front Pharmacol
January 2025
Center for Pharmacometrics and System Pharmacology, College of Pharmacy, University of Florida, Orlando, FL, United States.
Introduction: This study evaluated the relationship between total plasma and free kidney concentrations of amphotericin B (AmB) in healthy and -infected Wistar rats using microdialysis and has the potential to significantly impact future research in this field and promote the development of antifungal drugs. The findings of this study, which show that plasma levels are a good predictor for AmB kidney concentrations and can be used to optimize its dosing regimen, underscore the importance of this research.
Methods: Microdialysis probe recovery rates were determined by dialysis and retrodialysis , as well as by retrodialysis .
Interplay between energy metabolism and NADPH oxidase-mediated pathophysiology in cardiovascular diseases.
Front Pharmacol
January 2025
Department of Emergency Medicine, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan, China.
Sustained production of reactive oxygen species (ROS) and an imbalance in the antioxidant system have been implicated in the development of cardiovascular diseases (CVD), especially when combined with diabetes, hypercholesterolemia, and other metabolic disorders. Among them, NADPH oxidases (NOX), including NOX1-5, are major sources of ROS that mediate redox signaling in both physiological and pathological processes, including fibrosis, hypertrophy, and remodeling. Recent studies have demonstrated that mitochondria produce more proteins and energy in response to adverse stress, corresponding with an increase in superoxide radical anions.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!