Cytochrome P450 3A4 ELISA Kit (CYP3A4)

INTENDED USE

Human cytochrome P450 3A4 ELISA kit can measure concentrations of CYP3A4 (cytochrome P450 3A4, CYP3A3, Nifedipine, albendazole monooxygenase) present is plasma, serum, biological fluids or tissue homogenate samples. This assay has a minimum sensitivity detection limit of 46.875 pg/ml.

INTRODUCTION

Representing one of the most versatile and abundant drug metabolizing enzymes in humans, cytochrome P450 3A4 (CYP3A4) catalyzes oxidative biotransformation of over 50% of pharmaceuticals. Initial characterization began in the 1980s when cDNA isolation revealed a heme-containing monooxygenase selectively concentrated in human liver showing similarity to rat P450 forms. Early functional analyses uncovered potent abilities to metabolize widely used drugs including erythromycin, cyclosporine, and steroids. This exceptional catalytic diversity arises from CYP3A4’s large, flexible binding pocket, facilitating interaction with many structurally unrelated compounds.

However researchers soon discovered marked interindividual variability in CYP3A4 expression and activity – necessitating methods gauging function in patients to optimize drug dosing. Techniques quantifying altered drug metabolite ratios or clearance form a mainstay for assessing CYP3A4 capacity today. Another vital source of variability stems from induction or inhibition mediated by concomitant drugs, herbs, or endogenous compounds that dynamically alter CYP3A4 levels. For example, the antibiotic rifampin strongly upregulates hepatic CYP3A4 expression over 10-fold within days before homeostatic repression. Food constituents, inflammatory signals, and nuclear receptor signaling also tune CYP3A4-activity through transcriptional effects.

Moving forward, pharmacogenomic strategies seek to leverage knowledge of genetic polymorphisms contributing to expression differences to improve adverse event prediction and allow pretreatment screening. However, realization of true personalized medicine requires further illumination of the diverse exogenous and endogenous factors finely governing CYP3A4 at any moment. In particular, tissue-specific regulation by microRNAs, epigenetics, and splice variation represent intriguing areas undergoing active investigation. Overall the exceptional abundance and adaptability of CYP3A4 ensures its enduring importance customizing drug fate.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

• One 96-Well Plate: Pre-coated with anti- CYP3A4.
• Standards: Lyophilized recombinant.
• Sample/Standard Dilution Buffer.
• Biotinylated-labelled Antibody.
• Antibody Dilution Buffer.
• HRP-Streptavidin Conjugate (SABC).
• SABC Dilution Buffer.
• TMB Substrate.
• Wash Buffer (25x).
• Plate Sealer.
• Product Instructions.

TYPICAL RESULTS

For this CYP3A4 ELISA kit, it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 78.125, 156.25, 312.5, 625, 1250, 2500, 5000 pg/ml
Reactivity: Human
Sensitivity: 46.875 pg/ml
Range: 78.125 – 5000 pg/ml
Principle: Sandwich, Double Antibody
Application: Research Use Only.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for CYP3A4, no cross reactivity or interference between CYP3A4 and analogues was detected.
– Recovery: Serum (92 – 102%), EDTA Plasma (94 – 103%), Heparin Plasma (93 – 104%).
– Linearity: Serum (86 – 103%), EDTA Plasma (87 – 98%), Heparin Plasma (85 – 95%).
– Precison Intra-Assay: CV < 8%
– Precison Inter-Assay: CV < 10%

REFERENCES

1. Cytochrome P450 3A4 (CYP3A4) protein quantification using capillary western blot technology and total protein normalization. J Pharmacol Toxicol Methods. (2021) 112: 107117. Collins J.M. and Wang D.
2. Functional gene variants of CYP3A4. Clin Pharmacol Ther. (2014) 96 (3): 340-8. Werk A.N. and Cascorbi I.
3. Chimeric cytochrome P450 3A4 used for in vitro prediction of food-drug interactions. Biotechnol Appl Biochem. (2020) 67 (4): 541-548. Sadeghi S.J., et al.
4. Heterotropic cooperativity of cytochrome P450 3A4 and potential drug-drug interactions. Curr Drug Metab. (2001) 2 (2): 185-98. Tang W. and Stearns R.A.

ADDITIONAL INFORMATION

• Full Name: Cytochrome P450 3A4 ELISA Kit (CYP3A4)
• Reactivity: Human
• Sample Type: Plasma, Tissue Homogenates, Serum, Biological Fluids
• Sensitivity: 46.875 pg/ml

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